LOCUS Exported 3853 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of lambda-CI fused via three alanines to E. coli RNAP beta subunit, aa 831-1057 (beta-flap). Can be cut with Not1 and BamHI to replace beta-flap with a moiety of interest.. ACCESSION . VERSION . KEYWORDS pAC-lambda-CI-beta-flap (831-1057) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3853) AUTHORS Dove SL, Joung JK, Hochschild A TITLE Activation of prokaryotic transcription through arbitrary protein-protein contacts. JOURNAL Nature. 1997 Apr 10;386(6625):627-30. PUBMED 9121589 REFERENCE 2 (bases 1 to 3853) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3853 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 134..153 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(220..322) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" primer_bind complement(714..731) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(848..1392) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(928..947) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 1555..1585 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 1593..1609 /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 1598..1620 /label=M13/pUC Reverse /note="In lacZ gene" CDS 1630..2340 /codon_start=1 /gene="cI" /product="phage lambda repressor" /label=-lambda repressor /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV VGKVIASQWPEETFG" CDS complement(3413..219) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" ORIGIN 1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt 61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt 121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga 181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga 241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt 301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc 361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat 421 ttattcggcg caaagtgcgt cgggtgaatg ctgccaactt actgattagt gtatgatggt 481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg 541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact 601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa 661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc 721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc 781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa 841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc 901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttcccctg gcggctccct 961 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg 1021 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact 1081 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg 1141 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta 1201 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt 1261 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc 1321 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca 1381 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa 1441 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca 1501 tgtttgacag cttatcatcg ataagctaat tctcactcat taggcacccc aggctttaca 1561 ctttatgctt ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg 1621 aaacagcgta tgagcacaaa aaagaaacca ttaacacaag agcagcttga ggacgcacgt 1681 cgccttaaag caatttatga aaaaaagaaa aatgaacttg gcttatccca ggaatctgtc 1741 gcagacaaga tggggatggg gcagtcaggc gttggtgctt tatttaatgg catcaatgca 1801 ttaaatgctt ataacgccgc attgcttgca aaaattctca aagttagcgt tgaagaattt 1861 agcccttcaa tcgccagaga aatctacgag atgtatgaag cggttagtat gcagccgtca 1921 cttagaagtg agtatgagta ccctgttttt tctcatgttc aggcagggat gttctcacct 1981 gagcttagaa cctttaccaa aggtgatgcg gagagatggg taagcacaac caaaaaagcc 2041 agtgattctg cattctggct tgaggttgaa ggtaattcca tgaccgcacc aacaggctcc 2101 aagccaagct ttcctgacgg aatgttaatt ctcgttgacc ctgagcaggc tgttgagcca 2161 ggtgatttct gcatagccag acttgggggt gatgagttta ccttcaagaa actgatcagg 2221 gatagcggtc aggtgttttt acaaccacta aacccacagt acccaatgat cccatgcaat 2281 gagagttgtt ccgttgtggg gaaagttatc gctagtcagt ggcctgaaga gacgtttggc 2341 gcggccgcaa tccacattca ggaactggcg tgtgtgtccc gtgacaccaa gctgggtccg 2401 gaagagatca ccgctgacat cccgaacgtg ggtgaagctg cgctctccaa actggatgaa 2461 tccggtatcg tttacattgg tgcggaagtg accggtggcg acattctggt tggtaaggta 2521 acgccgaaag gtgaaactca gctgacccca gaagaaaaac tgctgcgtgc gatcttcggt 2581 gagaaagcct ctgacgttaa agactcttct ctgcgcgtac caaacggtgt atccggtacg 2641 gttatcgacg ttcaggtctt tactcgcgat ggcgtagaaa aagacaaacg tgcgctggaa 2701 atcgaagaaa tgcagctcaa acaggcgaag aaagacctgt ctgaagaact gcagatcctc 2761 gaagcgggtc tgttcagccg tatccgtgct gtgctggtag ccggtggcgt tgaagctgag 2821 aagctcgaca aactgccgcg cgatcgctgg ctggagctgg gcctgacaga cgaagagaaa 2881 caaaatcagc tggaacagct ggctgagcag tatgacgaac tgaaacacga gttcgagaag 2941 aaactcgaag cgaaacgccg caaaatcacc cagggcgacg atctggcacc gggcgtgctg 3001 aagattgtta aggtatatct ggcggttaaa taaggatcct taccttacag atctgcatcg 3061 caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctaaccgt 3121 ttttatcagg ctctgggagg cagaataaat gatcatatcg tcaattatta cctccacggg 3181 gagagcctga gcaaactggc ctcaggcatt tgagaagcac acggtcacac tgcttccggt 3241 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga 3301 accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt 3361 attcaggcgt agcaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc 3421 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc 3481 catcacagac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg 3541 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta 3601 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa 3661 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt 3721 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt 3781 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt 3841 tcattgccat acg //