LOCUS Exported 8394 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION GeneGuard genomic cassette: for constructing Delta-thyA::RBS B repA, Epsilon, FRT-kanR-FRT (low-copy support for ColE2 COR). Will support pSEVAT7Z.. ACCESSION . VERSION . KEYWORDS pSEVA12TEC-B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8394) AUTHORS Wright O, Delmans M, Stan GB, Ellis T TITLE GeneGuard: a Modular Plasmid System Designed for Biosafety. JOURNAL ACS Synth Biol. 2014 May 13. PUBMED 24847673 REFERENCE 2 (bases 1 to 8394) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8394 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(5..682) /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" promoter 1321..1355 /label=J23119 promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119)" terminator 2300..2371 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2387..2414 /label=T7Te terminator /note="phage T7 early transcription terminator" RBS 2483..2491 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" RBS 2531..2539 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" protein_bind 2834..2867 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" CDS 3235..4029 /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(3289..3308) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3899..3918 /label=Neo-F /note="Neomycin resistance gene, forward primer" protein_bind 4042..4089 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." terminator 4625..4719 /label=lambda t0 terminator /note="transcription terminator from phage lambda" promoter 4792..4883 /gene="bla" /label=AmpR promoter CDS 4884..5744 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" terminator 5748..5787 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" oriT 5934..6042 /note="incP origin of transfer" CDS complement(6103..7251) /codon_start=1 /product="trans-acting replication protein that binds to and activates oriV" /label=trfA /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" rep_origin 7644..8175 /label=oriV /note="incP origin of replication" terminator 8302..8388 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ttaattaagc accggtggag tgacgacctt cagcacgttc gtactgttca acgatggtgt 61 agtcttcgtt gtgggaggtg atgtccagtt tgatgtcggt tttgtaagca cccggcagct 121 gaaccggttt tttagccatg taggtggttt taacttcagc gtcgtagtga ccaccgtctt 181 tcagtttcag acgcattttg atttcacctt tcagagcacc gtcttccggg tacatacgtt 241 cggtggaagc ttcccaaccc atggtttttt tctgcataac cggaccgtcg gacgggaagt 301 tggtaccacg cagtttaact ttgtagatga actcaccgtc ttgcagggag gagtcctggg 361 taacggtaac aacaccaccg tcttcgaagt tcataacacg ttcccatttg aaaccttccg 421 ggaaggacag tttcaggtag tccgggatgt cagccgggtg tttaacgtaa gctttggaac 481 cgtactggaa ctgcggggac aggatgtccc aagcgaacgg cagcggacca cctttggtaa 541 ctttcagttt agcggtctgg gtaccttcgt acggacgacc ttcaccttca ccttcgattt 601 cgaactcgtg accgttaacg gaaccttcca tacgaacttt gaaacgcatg aactctttga 661 taacgtcttc ggaggaagcc atctagtatt caccaccttt ctctagtaac atttattgta 721 caacacgagc ccatttttgt caaataaaat ttaaattata tcaacgttaa taagacgttg 781 tcaataaaat tattttgaca aaattcctag gtctctgatt ttatcgcacc actcattccg 841 tttggtcttg gtgccgggcg tctgggcaac tttattaacg gtgaattgtg gggccgcgtt 901 gacccgaact tcccgtttgc catgctgttc cctggctccc gtacagaaga tattttgctg 961 ctgcaaacca acccgcagtg gcaatccatt ttcgacactt acggtgtgct gccgcgccac 1021 ccatcacagc tttacgagct gctgctggaa ggtgtggtgc tgtttattat cctcaacctg 1081 tatattcgta aaccacgccc aatgggagct gtctcaggtt tgttcctgat tggttacggc 1141 gcgtttcgca tcattgttga gtttttccgc cagcccgacg cgcagtttac cggtgcctgg 1201 gtgcagtaca tcagcatggg gcaaattctt tccatcccga tgattgtcgc gggtgtgatc 1261 atgatggtct gggcatatcg tcgcagccca cagcaacacg tttcctgagg aaccgaattc 1321 ttgacagcta gctcagtcct aggtataatg ctagcggatc tcgtcagtaa cgagccctgt 1381 gagaagtgaa cgtgagcgcc gtacttcaac gcttccgtga aaaattaccg cacaaaccgt 1441 actgtactaa cgatttcgcg tacggcgttc gcattctgcc gaaaaacatt gccattcttg 1501 cccgtttcat ccagcagaac cagccacatg cactgtactg gcttcccttt gacgtggacc 1561 ggacgggggc atcaatcgac tggagcgacc ggaattgtcc ggccccgaac atcaccgtaa 1621 aaaatccccg taacgggcac gcgcatctgc tctacgcgct cgcccttcct gtgagaactg 1681 cgccggatgc atcggcttcg gcgctcagat acgctgccgc tattgagcgt gcgttgtgtg 1741 aaaaactggg cgcggatgtg aattacagcg gcctgatctg caaaaatccg tgccaccctg 1801 aatggcagga agtggaatgg cgcgaggaac cctacactct cgacgaactg gctgattatc 1861 tggatctgag cgcctcagcg cgccgtagcg tcgataaaaa ttacgggctg gggcgaaact 1921 accatctgtt cgaaaaggtc cgtaaatggg cctacagagc gattcgtcag ggctggcctg 1981 tattctcaca atggcttgat gccgttatcc agcgtgtcga aatgtacaac gcttcgcttc 2041 ccgttccgct ttctccggct gaatgtcggg ctattggcaa gagcattgcg aaatatacac 2101 acaggaaatt ctcaccagag ggattttccg ctgtacaggc cgctcgcggt cgcaagggcg 2161 gaactaaatc taagcgcgca gcagttccta catcagcacg ttcgctgaaa ccgtgggagg 2221 cattaggcat cagtcgagcg acgtactacc gaaaattaaa atgtgaccca gacctcgcaa 2281 aataaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg 2341 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg 2401 tgggcctttc tgcgtttata cccgggaaaa agagtattga cttaaagtct aacctatagg 2461 tataatgtgt ggagaccaga ggtaaggagg taacaaccat gcgagtgttg aagaaacatc 2521 ttaatcatgc taaggaggtt ttctaatggc agttacgtat gaaaaaacat ttgaaataga 2581 gatcattaac gaattatcgg caagcgttta taatcgagta ttaaactatg ttttgaacca 2641 tgaattaaat aaaaatgact ctcaattatt ggaagtcaat ttattaaacc aattaaagct 2701 tgcaaaacgt gtaaatcttt ttgattattc tttagaagaa ttacaagccg ttcatgagta 2761 ttggcggtca atgaatcgtt actcaaaaca agttttgaat aaagagaaag tggcttgagt 2821 cgacctgcag ttcgaagttc ctattctcta gaaagtatag gaacttcaga gcgcttttga 2881 agctcacgct gccgcaagca ctcagggcgc aagggctgct aaaggaagcg gaacacgtag 2941 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg 3001 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga 3061 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc 3121 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc 3181 tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 3241 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3301 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 3361 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 3421 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 3481 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 3541 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 3601 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3661 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3721 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgtatgcc cgacggcgag 3781 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3841 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3901 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 3961 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4021 ttcttctaat aaggggatct tgaagttcct attccgaagt tcctattctc tagaaagtat 4081 aggaacttcg aagcagctcc agcctacagc atgcaaagcg ccggtggcta tctaattacg 4141 aaacatcctg ccagagccga cgccagtgtg cgtcggtttt tttaccctcc gttaaattct 4201 tcgagacgcc ttcccgaaat tttgcaacgt cctgcaacgg cgtaaatagt ccggaagatg 4261 cgccgaagaa atagaaactg gccctcgtct ttttttctcc tcgctccata ctgccggcat 4321 gaaaacacaa cgtggttata cgctgattga aacgctggtc gcgatgctga ttttggtcat 4381 gctaagcgca agtggactct atggctggca atactggcag cagtcgcaac ggctatggca 4441 aaccgccagc caggcgcggg actatttgct ctatttacgt gaagatgcca actggcataa 4501 ccgcgaccac agtatcagcg ttatcaggga ggggacgtta tggtgccttg tgagttccgc 4561 tgctggggcc aatacctgtc atggcagttc accattggtc tttgtgccac gctgactagt 4621 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga 4681 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc caataattac 4741 gatttaaatt agtagcccgc ctaatgagcg ggcttttttt taattcccct atttgtttat 4801 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 4861 aataatattg aaaaaggaag agtatgagca ttcagcattt tcgtgtggcg ctgattccgt 4921 tttttgcggc gttttgcctg ccggtgtttg cgcatccgga aaccctggtg aaagtgaaag 4981 atgcggaaga tcaactgggt gcgcgcgtgg gctatattga actggatctg aacagcggca 5041 aaattctgga atcttttcgt ccggaagaac gttttccgat gatgagcacc tttaaagtgc 5101 tgctgtgcgg tgcggttctg agccgtgtgg atgcgggcca ggaacaactg ggccgtcgta 5161 ttcattatag ccagaacgat ctggtggaat atagcccggt gaccgaaaaa catctgaccg 5221 atggcatgac cgtgcgtgaa ctgtgcagcg cggcgattac catgagcgat aacaccgcgg 5281 cgaacctgct gctgacgacc attggcggtc cgaaagaact gaccgcgttt ctgcataaca 5341 tgggcgatca tgtgacccgt ctggatcgtt gggaaccgga actgaacgaa gcgattccga 5401 acgatgaacg tgataccacc atgccggcag caatggcgac caccctgcgt aaactgctga 5461 cgggtgagct gctgaccctg gcaagccgcc agcaactgat tgattggatg gaagcggata 5521 aagtggcggg tccgctgctg cgtagcgcgc tgccggctgg ctggtttatt gcggataaaa 5581 gcggtgcggg cgaacgtggc agccgtggca ttattgcggc gctgggcccg gatggtaaac 5641 cgagccgtat tgtggtgatt tataccaccg gcagccaggc gacgatggat gaacgtaacc 5701 gtcagattgc ggaaattggc gcgagcctga ttaaacattg gtaaaccgat acaattaaag 5761 gctccttttg gagccttttt ttttggacga cccttgtcct tttccgctgc ataaccctgc 5821 ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg atatacagga 5881 ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca gccgggcagg 5941 ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc ttattcgcac 6001 ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgtaggccgg ccgcgatgca 6061 ggtggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac 6121 caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc 6181 ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg 6241 gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg 6301 tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc 6361 agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc 6421 acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc 6481 ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta 6541 ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat 6601 cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc 6661 atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa 6721 aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc 6781 agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa 6841 caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt 6901 ggcttcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat 6961 agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcgagacg 7021 acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct 7081 gtcgcgctcg atcttggccg tagcttgctg gactatcgag ccgacggact ggaaggtttc 7141 gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc 7201 cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc 7261 ttgcgggatt gccccggaat taattccccg gatcgatccg tcgatcttga tcccctgcgc 7321 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta 7381 ccagagggcg ccccagctgg caattccggt tcgcttgctg tccataaaac cgcccagtct 7441 agctatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct tgcgttttcc 7501 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg 7561 gctttctacg tggctgccat ttttggggtg aggccgttcg cggccgaggg gcgcagcccc 7621 tggggggatg ggaggcccgc gttagcgggc cgggagggtt cgagaagggg gggcaccccc 7681 cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg ttaaaaacaa ggtttataaa 7741 tattggttta aaagcaggtt aaaagacagg ttagcggtgg ccgaaaaacg ggcggaaacc 7801 cttgcaaatg ctggattttc tgcctgtgga cagcccctca aatgtcaata ggtgcgcccc 7861 tcatctgtca gcactctgcc cctcaagtgt caaggatcgc gcccctcatc tgtcagtagt 7921 cgcgcccctc aagtgtcaat accgcagggc acttatcccc aggcttgtcc acatcatctg 7981 tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc gaggctggcc agctccacgt 8041 cgccggccga aatcgagcct gcccctcatc tgtcaacgcc gcgccgggtg agtcggcccc 8101 tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc aagttttccg cgaggtatcc 8161 acaacgccgg cggccctaca tggctctgct gtagtgagtg ggttgcgctc cggcagcggt 8221 cctgatcccc cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggcgcg 8281 cccagctgtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca 8341 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcct //