LOCUS Exported 5391 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cloning vector containing A-B-C casette with linker L3 in site B. ACCESSION . VERSION . KEYWORDS pOVC1_L3(GSSSSGGSSSG) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5391) 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 5391) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5391 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 158..537 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 538..741 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 692..712 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 786..805 /label=T7 /note="T7 promoter, forward primer" promoter 786..804 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(905..922) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 911..1135 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1181..1609 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1268..1287) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1478..1499 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1618..1638) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1623..1952 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1803..1938 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1865..1884 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2019..2813 /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(2073..2092) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2683..2702 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 2987..3108 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3024..3043) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3078..3097 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3157..3173) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3157..3173) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3170..3192) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3181..3197 /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(3205..3235) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3250..3271 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3388..3405) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3559..4147) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3639..3658) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4318..5178) /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 4941..4960 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5179..5283) /gene="bla" /label=AmpR promoter primer_bind complement(5358..5377) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct 61 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt 121 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat 181 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 541 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 601 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 661 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 721 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc 781 gaaattaata cgactcacta tagggagacc caagctgacc atggccaccg gtgaattcgg 841 aagcagctct tctggcggat ccagtagtgg tggatccccc gggtaagttt aaaccgctga 901 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 961 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 1021 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 1081 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 1141 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 1201 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 1261 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 1321 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 1381 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 1441 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 1501 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 1561 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 1621 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 1681 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 1741 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 1801 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 1861 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 1921 gaggcttttt tggaggccta ggcttttgca aaaagctctc gggagcttgt atatccattt 1981 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 2041 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 2101 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 2161 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 2221 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 2281 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 2341 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 2401 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 2461 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 2521 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 2581 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 2641 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 2701 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 2761 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 2821 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 2881 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 2941 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc 3001 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 3061 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 3121 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 3181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 3241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 3301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 3361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 3421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 3481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 3541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 3601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 3661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 3721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 3781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 3841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 3901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 3961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 4021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 4261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 4321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 4381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 4441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 4501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 4561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 4621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 4681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 4741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 4801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 4861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 4921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 4981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 5041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 5101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 5161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 5221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 5281 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc 5341 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca g //