LOCUS Exported 5376 bp DNA circular SYN 02-JUL-2021 DEFINITION BBF10K_000516 in pOpen_v4. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5376) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 2, 2021 from SnapGene 5.3.1 https://www.snapgene.com COMMENT This file was created using tools provided by Twist Bioscience FEATURES Location/Qualifiers source 1..5376 /organism="BBF10K_000516 in pOpen_v4" primer_bind 21..51 /label=Gibson-for regulatory 46..55 /regulatory_class="other" /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 52..882 /codon_start=1 /product="cluster of differentiation 40" /label=CD40 /translation="MVRLPLQCVLWGCLLTAVHPEPPTACREKQYLINSQCCSLCQPGQ KLVSDCTEFTETECLPCGESEFLDTWNRETHCHQHKYCDPNLGLRVQQKGTSETDTICT CEEGWHCTSEACESCVLHRSCSPGFGVKQIATGVSDTICEPCPVGFFSNVSSAFEKCHP WTSCETKDLVVQQAGTNKTDVVCGPQDRLRALVVIPIIFGILFAILLVLVFIKKVAKKP TNKAPHPKQEPQEINFPDDLPGSNTAAPVQETLHGCQPVTQEDGKESRISVQERQ" primer_bind complement(883..907) /label=Gibson-rev terminator 1026..1073 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(1078..1094) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" misc_feature 1253..1503 /gene="incC" /label=incC /note="incompatibility region of the bacterial F plasmid" CDS complement(1506..2261) /codon_start=1 /gene="repE" /product="replication initiation protein for the bacterial F plasmid" /label=repE /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM RLSYIEKKKGRQTTHIVFSFRDITSMTTG" rep_origin 2352..2571 /label=ori2 /note="secondary origin of replication for the bacterial F plasmid; also known as oriS" protein_bind 2991..3012 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3027..3057 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3065..3081 /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 3089..3105 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS 3178..4023 /codon_start=1 /label=repL /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG " misc_feature 4131..4282 /label=nnpp promoters CDS 4322..5137 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" terminator complement(5247..5290) /label=bacterial terminator /note="putative bacterial transcription terminator" primer_bind 5308..5324 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" terminator 5325..5356 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catggtgaga 61 ctgcccctgc agtgcgtgct gtggggctgc ctgctgaccg ccgtgcaccc cgagcccccc 121 accgcctgca gagagaagca gtacctgatc aacagccagt gctgcagcct gtgccagccc 181 ggccagaagc tggtgagcga ctgcaccgag ttcaccgaga ccgagtgcct gccctgcggc 241 gagagcgagt tcctggacac ctggaacaga gagacccact gccaccagca caagtactgc 301 gaccccaacc tgggcctgag agtgcagcag aagggcacca gcgagaccga caccatctgc 361 acctgcgagg agggctggca ctgcaccagc gaggcctgcg agagctgcgt gctgcacaga 421 agctgcagcc ccggcttcgg cgtgaagcag atcgccaccg gcgtgagcga caccatctgc 481 gagccctgcc ccgtgggctt cttcagcaac gtgagcagcg ccttcgagaa gtgccacccc 541 tggaccagct gcgagaccaa ggacctggtg gtgcagcagg ccggcaccaa caagaccgac 601 gtggtgtgcg gcccccagga cagactgaga gccctggtgg tgatccccat catcttcggc 661 atcctgttcg ccatcctgct ggtgctggtg ttcatcaaga aggtggccaa gaagcccacc 721 aacaaggccc cccaccccaa gcaggagccc caggagatca acttccccga cgacctgccc 781 ggcagcaaca ccgccgcccc cgtgcaggag accctgcacg gctgccagcc cgtgacccag 841 gaggacggca aggagagcag aatcagcgtg caggagagac agggatccgc caccaacttc 901 agcctgctag atagtcacgc acaagaccac cctttgcggg ctgactgact agggataaca 961 gggtaatgct ctatttcatg tccaacaccg ctgaggtgtc aatcgtcgga gccgctgagc 1021 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgcatggtc 1081 atagctgttt cctgagagct tggcaggtgt cgcgtatgca tcgatcacgt gtccacgttc 1141 tttaatagtg gactcttgtt ccaaactgga acaacactcg gatcgatccg gcgcgcaccg 1201 tgggaaaaac tccaggtaga ggtacacacg cggatagcca attcagagta ataaactgtg 1261 ataatcaacc ctcatcaatg atgacgaact aacccccgat atcaggtcac atgacgaagg 1321 gaaagagaag gaaatcaact gtgacaaact gccctcaaat ttggcttcct taaaaattac 1381 agttcaaaaa gtatgagaaa atccatgcag gctgaaggaa acagcaaaac tgtgacaaat 1441 taccctcagt aggtcagaac aaatgtgacg aaccaccctc aaatctgtga cagataaccc 1501 tcagactatc ctgtcgtcat ggaagtgata tcgcggaagg aaaatacgat atgagtcgtc 1561 tggcggcctt tctttttctc aatgtatgag aggcgcattg gagttctgct gttgatctca 1621 ttaacacaga cttgcaggaa gcggcggcgg aagtcaggca tacgctggta actttgaggc 1681 agctggtaac gctctatgat ccagtcgatt ttcagagaca cgatgcctga gccatccggc 1741 ttacgatact gacacaggga ttcgtataaa cgcatggcat acggattggt gatttctttt 1801 gtttcactaa gccgaaactg cgtaaaccgg ttctgtaacc cgataaagaa gggaatgaga 1861 tatgggttga tatgtacact gtaaagccct ctggatggac tgtgcgcacg tttgataaac 1921 caaggaaaag attcatagcc tttttcatca ccggcatcct cttcagggcg ataaaaaacc 1981 acttccttcc ccgcgaaact cttcaatgcc tgccgtatat ccttactggc ttccgcagag 2041 gtcaatccga atatttcagc atatttagca acatggatct cgcagatacc gtcatgttcc 2101 tgtagggtgc catcagattt tctgatctgg tcaacgaaca gatacagcat acgtttttga 2161 tcccgggaga gactatatgc cgcctcagtg aggtcgtttg actggacgat tcgcgggcta 2221 tttttacgtt tcttgtgatt gataaccgct gtttccgcca tgacagatcc atgtgaagtg 2281 tgacaagttt ttagattgtc acactaaata aaaaagagtc aataagcagg gataactttg 2341 tgaaaaaaca gcttcttctg agggcaattt gtcacagggt taagggcaat ttgtcacaga 2401 caggactgtc atttgagggt gatttgtcac actgaaaggg caatttgtca caacaccttc 2461 tgtagaacca gcatggataa aggcctacaa ggcgctctaa aaaagaagat ctaaaaacta 2521 taaaaaaaat aattataaaa atatccccgt ggataagtgg ataaccccaa gggaagtttt 2581 ttcaggcatc gtgtgtaagc agaatatata agtgctgttc cctggtgctt cctcgctcac 2641 tcgaaattcc cggggatagc tttatgcttg taaaccgttt tgtgaaaaaa tttttaaaat 2701 aaaaaagggg acctctaggg tccccaatta attagtaata taatctatta aaggtcattc 2761 aaaaggtcat ccaccggatc aattcccctg ctcgcgcagg ctgggtgcca agctctcggg 2821 taacatcaag gcccgatcct tggagccctt gccctcccgc acgatgatcg tgccgtgatc 2881 gaaatccaga tccttgaccc gcagttgcaa accctcactg atccgattca ttaatgcagc 2941 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt 3001 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt 3061 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag 3121 cgcggtacct gtcgcggcaa cgcgctaaca gacgtagtaa gaaccaccag cattgtaatg 3181 ctggctaaag tcactttcct gagctgtata acgatgagcg attttacttt ttctggctat 3241 gaattggcct gctttgtaac acactccggt ctatcccgta gcgccgggca tatcctgtcg 3301 caatgtgcaa atctcgcggc aacaaccagt gaatacttca ttcacaagcc tcaccgcctg 3361 atcgcggcag aaactggtta tagccaatca accgtcgttc gtgcattccg tgaagctgta 3421 aacaaaggta tcctgtctgt agagattgtt atcggcgatc accgtgaacg tcgcgctaac 3481 ctgtaccggt ttacaccatc ctttttggcc ttcgcacaac aagccaaaaa tgcgctgata 3541 gaaagcaaat taaagatctc ttcagcggca accaaggtta aagctgttct cgctaagaca 3601 ttggctttat ttaatttttt atccacaccc ccatgtcaaa atgatacccc ctccccctgt 3661 caggatgacg tggcaataaa gaataagaag tcacaagtta aaaaaacaaa aagatcagtt 3721 tccggcggtg ccggaacaac cagcctcaaa aaattgactt catggatcgc taaggcaaaa 3781 gcaaaggctg acaatctgcg gttatccaaa aaacgcactc aaaaacatga gttcaagcag 3841 aaagtagagg cggctgcgcg gaaatatgct tacctgaaga acaagcgttc gcctgatatt 3901 ggcgggatat caaacttcga taacctaccg cattgcatga cggtaaacga agctcttaat 3961 gcggttttag ccaaaaataa agataacgaa caatggggta taccggcagg attcagaggg 4021 taatgaattg ctctaattat aaccatgcat actttcaaca cctctagttt gccatgaggc 4081 aaactcatag gtgtcctggt aagaggacac tgttgccaaa actggacgcc ccattattgc 4141 aattaataaa caactaacgg acaattctac ctaacaataa gtggagttgc ggccggccgc 4201 gatccgctag caaagccacg ttgtgtctca aaatctctga tgttacattg cacaagataa 4261 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt 4321 tatgagccat attcaacggg aaacgtcttg ctcgcgcccg cgattaaatt ccaacatgga 4381 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat 4441 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag 4501 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc 4561 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc 4621 gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat 4681 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc 4741 ttttaacagc gatcgcgtat ttcgcctcgc tcaggcgcaa tcacgaatga ataacggttt 4801 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa 4861 agaaatgcat aaactgttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc 4921 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt 4981 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc 5041 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa 5101 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta 5161 acactggcga gctctgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 5221 taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa 5281 ggatgatttc tgcacctgca ccagtcagta aaacgacggc cagtagtcaa aagcctccga 5341 ccggaggctt ttgacttggt tcaggtggag tgggag //