LOCUS Exported 8484 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION High core fragment expression without SynZIP. ACCESSION . VERSION . KEYWORDS pTHSSd_40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8484) AUTHORS Segall-Shapiro TH, Meyer AJ, Ellington AD, Sontag ED, Voigt CA TITLE A 'resource allocator' for transcription based on a highly fragmented T7 RNA polymerase. JOURNAL Mol Syst Biol. 2014 Jul 30;10(7):742. PUBMED 25080493 REFERENCE 2 (bases 1 to 8484) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8484 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1807..1827 /label=BioBrick suffix /note="universal suffix for all parts" terminator 1863..1894 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" CDS 1980..1997 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2053..2070) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 2223..2269 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin 2325..2713 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" CDS complement(2763..3557) /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 SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind 3484..3503 /label=Neo-R /note="Neomycin resistance gene, reverse primer" rep_origin 3963..4182 /label=ori2 /note="secondary origin of replication for the bacterial F plasmid; also known as oriS" CDS 4273..5028 /codon_start=1 /gene="repE" /product="replication initiation protein for the bacterial F plasmid" /label=repE /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM RLSYIEKKKGRQTTHIVFSFRDITSMTTG" misc_feature 5031..5281 /gene="incC" /label=incC /note="incompatibility region of the bacterial F plasmid" CDS 5607..6782 /codon_start=1 /gene="sopA" /product="partitioning protein for the bacterial F plasmid" /label=sopA /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR" CDS 6782..7753 /codon_start=1 /gene="sopB" /product="partitioning protein for the bacterial F plasmid" /label=sopB /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP" misc_feature 7826..8299 /gene="sopC" /label=sopC /note="centromere-like partitioning region of the bacterial F plasmid" ORIGIN 1 atgaacacga ttaacatcgc taagaacgac ttctctgaca tcgaactggc tgctatcccg 61 ttcaacactc tggctgacca ttacggtgag cgtttagctc gcgaacagtt ggcccttgag 121 catgagtctt acgagatggg tgaagcacgc ttccgcaaga tgtttgagcg tcaacttaaa 181 gctggtgagg ttgcggataa cgctgccgcc aagcctctca tcactaccct actccctaag 241 atgattgcac gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg caagcgcccg 301 acagccttcc agttcctgca agaaatcaag ccggaagccg tagcgtacat caccattaag 361 accactctgg cttgcctaac cagtgctgac aatacaaccg ttcaggctgt agcaagcgca 421 atcggtcggg ccattgagga cgaggctcgc ttcggtcgta tccgtgacct tgaagctaag 481 cacttcaaga aaaacgttga ggaacaactc aacaagcgcg tagggcacgt ctacaagaaa 541 gcatttatgc aagttgtcga ggctgacatg ctctctaagg gtctactcgg tggcgaggcg 601 tggtcttcgt ggcataagga agactctatt catgtaggag tacgctgcat cgagatgctc 661 attgagtcaa ccggaatggt tagcttacac cgccaaaatg ctggcgtagt aggtcaagac 721 tctgagacta tcgaactcgc acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg 781 ctggctggca tctctccgat gttccaacct tgcgtagttc ctcctaagcc gtggactggc 841 attactggtg gtggctattg ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac 901 agtaagaaag cactgatgcg ctacgaagac gtttacatgc ctgaggtgta caaagcgatt 961 aacattgcgc aaaacaccgc atggaaaatc aacaagaaag tcctagcggt cgccaacgta 1021 atcaccaagt ggaagcattg tccggtcgag gacatccctg cgattgagcg tgaagaactc 1081 ccgatgaaac cggaagacat cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct 1141 gccgctgctg tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag ccttgagttc 1201 atgcttgagc aagccaataa gtttgctaac cataaggcca tctggttccc ttacaacatg 1261 gactggcgcg gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc 1321 aaaggactgc ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta ctactggctg 1381 aaaatccacg gtgcaaactg tgcgggtgtc gacaaggttc cgttccctga gcgcatcaag 1441 ttcattgagg aaaaccacga gaacatcatg gcttgcgcta agtctccact ggagaacact 1501 tggtgggctg agcaagattc tccgttctgc ttccttgcgt tctgctttga gtacgctggg 1561 gtacagcacc acggcctgag ctataactgc tcccttccgc tggcgtttga cgggtcttgc 1621 tctggcatcc agcacttctc cgcgatgctc cgagatgagg taggtggtcg cgcggttaac 1681 ttgcttccta gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa agtcaacgag 1741 attctacaag cagacgcaat caatgggacc gataacgaag tagttaccgt gaccgatgag 1801 aactaatact agtagcggcc gctgcaggag tcactaaggg ttagttagtt agattagcag 1861 aaagtcaaaa gcctccgacc ggaggctttt gactaaaact tcccttgggg ttatcattgg 1921 ggctcactca aaggcggtaa taactcatct cagaagagga tctgaatagc gccgtcgacc 1981 atcatcatca tcatcattga gtttaaacgg ccgccaaaac agccaagctg gagaatgaga 2041 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa 2101 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa 2161 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc 2221 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2281 cggtgaacta attctgattc gcacgggccc atggctaatt cccatgtcag ccgttaagtg 2341 ttcctgtgtc actcaaaatt gctttgagag gctctaaggg cttctcagtg cgttacatcc 2401 ctggcttgtt gtccacaacc gttaaacctt aaaagcttta aaagccttat atattctttt 2461 ttttcttata aaacttaaaa ccttagaggc tatttaagtt gctgatttat attaatttta 2521 ttgttcaaac atgagagctt agtacgtgaa acatgagagc ttagtacgtt agccatgaga 2581 gcttagtacg ttagccatga gggtttagtt cgttaaacat gagagcttag tacgttaaac 2641 atgagagctt agtacgtgaa acatgagagc ttagtacgta ctatcaacag gttgaactgc 2701 tgatcttcag atcctctacg ccggacgcat cgtggccgga tcttgcgcaa gcggtctcca 2761 attcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg 2821 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca 2881 cgggtatcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgttg 2941 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatggatc 3001 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc 3061 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga 3121 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca 3181 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg 3241 cgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct 3301 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc 3361 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga 3421 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt 3481 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat 3541 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc 3601 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca 3661 accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc 3721 cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg 3781 ttttcccttg tccagatagc ccagtagctg acattcatca agcttatgat gtggtctgtc 3841 cttttacagc cagtagtgct cgccgcagtc gagcgacagg gcgaagccct cgagtgagcg 3901 aggaagcacc agggaacagc acttatatat tctgcttaca cacgatgcct gaaaaaactt 3961 cccttggggt tatccactta tccacgggga tatttttata attatttttt ttatagtttt 4021 tagatcttct tttttagagc gccttgtagg cctttatcca tgctggttct agagaaggtg 4081 ttgtgacaaa ttgccctttc agtgtgacaa atcaccctca aatgacagtc ctgtctgtga 4141 caaattgccc ttaaccctgt gacaaattgc cctcagaaga agctgttttt tcacaaagtt 4201 atccctgctt attgactctt ttttatttag tgtgacaatc taaaaacttg tcacacttca 4261 catggatctg tcatggcgga aacagcggtt atcaatcaca agaaacgtaa aaatagcccg 4321 cgaatcgtcc agtcaaacga cctcactgag gcggcatata gtctctcccg ggatcaaaaa 4381 cgtatgctgt atctgttcgt tgaccagatc agaaaatctg atggcaccct acaggaacat 4441 gacggtatct gcgagatcca tgttgctaaa tatgctgaaa tattcggatt gacctctgcg 4501 gaagccagta aggatatacg gcaggcattg aagagtttcg cggggaagga agtggttttt 4561 tatcgccctg aagaggatgc cggcgatgaa aaaggctatg aatcttttcc ttggtttatc 4621 aaacgtgcgc acagtccatc cagagggctt tacagtgtac atatcaaccc atatctcatt 4681 cccttcttta tcgggttaca gaaccggttt acgcagtttc ggcttagtga aacaaaagaa 4741 atcaccaatc cgtatgccat gcgtttatac gaatccctgt gtcagtatcg taagccggat 4801 ggctcaggca tcgtctctct gaaaatcgac tggatcatag agcgttacca gctgcctcaa 4861 agttaccagc gtatgcctga cttccgccgc cgcttcctgc aggtctgtgt taatgagatc 4921 aacagcagaa ctccaatgcg cctctcatac attgagaaaa agaaaggccg ccagacgact 4981 catatcgtat tttccttccg cgatatcact tccatgacga caggatagtc tgagggttat 5041 ctgtcacaga tttgagggtg gttcgtcaca tttgttctga cctactgagg gtaatttgtc 5101 acagttttgc tgtttccttc agcctgcatg gattttctca tactttttga actgtaattt 5161 ttaaggaagc caaatttgag ggcagtttgt cacagttgat ttccttctct ttcccttcgt 5221 catgtgacct gatatcgggg gttagttcgt catcattgat gagggttgat tatcacagtt 5281 tattactctg aattggctat ccgcgtgtgt acctctacct ggagtttttc ccacggtgga 5341 tatttcttct tgcgctgagc gtaagagcta tctgacagaa cagttcttct ttgcttcctc 5401 gccagttcgc tcgctatgct cggttacacg gctgcggcga gcgctagtga taataagtga 5461 ctgaggtatg tgctcttctt atctcctttt gtagtgttgc tcttatttta aacaactttg 5521 cggttttttg atgactttgc gattttgttg ttgctttgca gtaaattgca agatttaata 5581 aaaaaacgca aagcaatgat taaaggatgt tcagaatgaa actcatggaa acacttaacc 5641 agtgcataaa cgctggtcat gaaatgacga aggctatcgc cattgcacag tttaatgatg 5701 acagcccgga agcgaggaaa ataacccggc gctggagaat aggtgaagca gcggatttag 5761 ttggggtttc ttctcaggct atcagagatg ccgagaaagc agggcgacta ccgcacccgg 5821 atatggaaat tcgaggacgg gttgagcaac gtgttggtta tacaattgaa caaattaatc 5881 atatgcgtga tgtgtttggt acgcgattgc gacgtgctga agacgtattt ccaccggtga 5941 tcggggttgc tgcccataaa ggtggcgttt acaaaacctc agtttctgtt catcttgctc 6001 aggatctggc tctgaagggg ctacgtgttt tgctcgtgga aggtaacgac ccccagggaa 6061 cagcctcaat gtatcacgga tgggtaccag atcttcatat tcatgcagaa gacactctcc 6121 tgcctttcta tcttggggaa aaggacgatg tcacttatgc aataaagccc acttgctggc 6181 cggggcttga cattattcct tcctgtctgg ctctgcaccg tattgaaact gagttaatgg 6241 gcaaatttga tgaaggtaaa ctgcccaccg atccacacct gatgctccga ctggccattg 6301 aaactgttgc tcatgactat gatgtcatag ttattgacag cgcgcctaac ctgggtatcg 6361 gcacgattaa tgtcgtatgt gctgctgatg tgctgattgt tcccacgcct gctgagttgt 6421 ttgactacac ctccgcactg cagtttttcg atatgcttcg tgatctgctc aagaacgttg 6481 atcttaaagg gttcgagcct gatgtacgta ttttgcttac caaatacagc aatagtaatg 6541 gctctcagtc cccgtggatg gaggagcaaa ttcgggatgc ctggggaagc atggttctaa 6601 aaaatgttgt acgtgaaacg gatgaagttg gtaaaggtca gatccggatg agaactgttt 6661 ttgaacaggc cattgatcaa cgctcttcaa ctggtgcctg gagaaatgct ctttctattt 6721 gggaacctgt ctgcaatgaa attttcgatc gtctgattaa accacgctgg gagattagat 6781 aatgaagcgt gcgcctgtta ttccaaaaca tacgctcaat actcaaccgg ttgaagatac 6841 ttcgttatcg acaccagctg ccccgatggt ggattcgtta attgcgcgcg taggagtaat 6901 ggctcgcggt aatgccatta ctttgcctgt atgtggtcgg gatgtgaagt ttactcttga 6961 agtgctccgg ggtgatagtg ttgagaagac ctctcgggta tggtcaggta atgaacgtga 7021 ccaggagctg cttactgagg acgcactgga tgatctcatc ccttcttttc tactgactgg 7081 tcaacagaca ccggcgttcg gtcgaagagt atctggtgtc atagaaattg ccgatgggag 7141 tcgccgtcgt aaagctgctg cacttaccga aagtgattat cgtgttctgg ttggcgagct 7201 ggatgatgag cagatggctg cattatccag attgggtaac gattatcgcc caacaagtgc 7261 ttatgaacgt ggtcagcgtt atgcaagccg attgcagaat gaatttgctg gaaatatttc 7321 tgcgctggct gatgcggaaa atatttcacg taagattatt acccgctgta tcaacaccgc 7381 caaattgcct aaatcagttg ttgctctttt ttctcacccc ggtgaactat ctgcccggtc 7441 aggtgatgca cttcaaaaag cctttacaga taaagaggaa ttacttaagc agcaggcatc 7501 taaccttcat gagcagaaaa aagctggggt gatatttgaa gctgaagaag ttatcactct 7561 tttaacttct gtgcttaaaa cgtcatctgc atcaagaact agcttaagct cacgacatca 7621 gtttgctcct ggagcgacag tattgtataa gggcgataaa atggtgctta acctggacag 7681 gtctcgtgtt ccaactgagt gtatagagaa aattgaggcc attcttaagg aacttgaaaa 7741 gccagcaccc tgatgcgacc acgttttagt ctacgtttat ctgtctttac ttaatgtcct 7801 ttgttacagg ccagaaagca taactggcct gaatattctc tctgggccca ctgttccact 7861 tgtatcgtcg gtctgataat cagactggga ccacggtccc actcgtatcg tcggtctgat 7921 tattagtctg ggaccacggt cccactcgta tcgtcggtct gattattagt ctgggaccac 7981 ggtcccactc gtatcgtcgg tctgataatc agactgggac cacggtccca ctcgtatcgt 8041 cggtctgatt attagtctgg gaccatggtc ccactcgtat cgtcggtctg attattagtc 8101 tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctggaacc acggtcccac 8161 tcgtatcgtc ggtctgatta ttagtctggg accacggtcc cactcgtatc gtcggtctga 8221 ttattagtct gggaccacga tcccactcgt gttgtcggtc tgattatcgg tctgggacca 8281 cggtcccact tgtattgtcg atcagactat cagcgtgaga ctacgattcc atcaatgcct 8341 gtcaagggca agtattgaca tgtcgtcgta acctgtagaa cggagtaacc tcggtgtgcg 8401 gttgtatgcc tgctgtggat tgtttacggc tagctcagtc ctaggtacta tgctagctac 8461 tagagtcatt tatgaaagta ctag //