LOCUS       sequence-89817-e        8156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GeneGuard genomic cassette: for constructing Delta-dapA::RBS E pi, 
            Kis, FRT-kanR-FRT (medium-copy support for R6K COR). Will support 
            pSEVAD1K..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1315 to 1349)
  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 8156)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2014 May 13."
COMMENT     Sequence Label: pSEVA12DKR-E
FEATURES             Location/Qualifiers
     source          1..8156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        1315..1349
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     terminator      2301..2372
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2388..2415
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2484..2492
                     /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             2532..2540
                     /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    2820..2853
                     /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             3221..4015
                     /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(3275..3294)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3885..3904
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    4028..4075
                     /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      4387..4481
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        4554..4645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4646..4714,4715..5506)
                     /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      5510..5549
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            5696..5804
                     /note="incP origin of transfer"
     CDS             complement(5865..7013)
                     /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      7406..7937
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(8064..8150)
                     /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 gcgtcgaaaa gtgctgtgaa gcgttcaatt
      841 aaatgcgggg agtctgccac atcgacctga acccagacag atgctggcat tggcggacgc
      901 ggacgcgccg tcgtgcgctt catcacgatt aaaagatcca gttcggcacc tttcaacggt
      961 aacgttgatt caatcagagt aatggcattc catgaaccgg aaagcagcat aataaacgtg
     1021 aactcttctc ccagcatcgc caggcgactg tcttcaatat tacagccgca actactgaca
     1081 tgacgggtga tggtgttcac aattccaggg cgatcggcac ccaacgcagt gatcaccaga
     1141 taatgttgcg atgacagtgt caaactggtt attcctttaa ggggtgagtt gttcttaagg
     1201 aaagcataaa aaaaacatgc atacaacaat cagaacggtt ctgtctgctt gcttttaatg
     1261 ccataccaaa cgtaccattg agacacttgt ttgcacagag gatggcccga attcttgacg
     1321 gctagctcag tcctaggtac agtgctagcg gatcttccgg aagactaggt gagactcaag
     1381 gtcatgatgg acgtgaacaa aaaaacgaaa attcgccacc gaaacgagct aaatcacacc
     1441 ctggctcaac ttcctttgcc cgcaaagcga gtgatgtata tggcgcttgc tcccattgat
     1501 agcaaagaac ctcttgaacg agggcgagtt ttcaaaatta gggctgaaga ccttgcagcg
     1561 ctcgccaaaa tcaccccatc gcttgcttat cgacaattaa aagagggtgg taaattactt
     1621 ggtgccagca aaatttcgct aagaggggat gatatcattg ctttagctaa agagcttaac
     1681 ctgctcttta ctgctaaaaa ctcccctgaa gagttagacc ttaacattat tgagtggata
     1741 gcttattcaa atgatgaagg atacttgtct ttaaaattca ccagaaccat agaaccatat
     1801 atctctagcc ttattgggaa aaaaaataaa ttcacaacgc aattgttaac ggcaagctta
     1861 cgcttaagta gccagtattc atcttctctt tatcaactta tcaggaagca ttactctaat
     1921 tttaagaaga aaaattattt tattatttcc gttgatgagt taaaggaaga gttaatagct
     1981 tatacttttg ataaagatgg aaatattgag tacaaatacc ctgactttcc tatttttaaa
     2041 agggatgtgt taaataaagc cattgctgaa attaaaaaga aaacagaaat atcgtttgtt
     2101 ggcttcactg ttcatgaaaa agaaggaaga aaaattagta agctgaagtt cgaatttgtc
     2161 gttgatgaag atgaattttc tggcgataaa gatgatgaag ctttttttat gaatttatct
     2221 gaagctgatg cagcttttct caaggtattt gatgaaaccg tacctcccaa aaaagctaag
     2281 gggtgaggat ctccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     2341 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
     2401 gtgggccttt ctgcgtttat acccgggaaa aagagtattg acttaaagtc taacctatag
     2461 gtataatgtg tggagaccag aggtaaggag gtaacaacca tgcgagtgtt gaagaaacat
     2521 cttaatcatg ctaaggaggt tttctaatgc ataccacccg actgaagagg gttggcggct
     2581 cagttatgct gaccgtccca ccggcactgc tgaatgcgct gtctctgggc acagataatg
     2641 aagttggcat ggtcattgat aatggccggc tgattgttga gccgtacaga cgcccgcaat
     2701 attcactggc tgagctactg gcacagtgtg atccgaatgc tgaaatatca gctgaagaac
     2761 gagaatggct ggatgcaccg gcgactggtc aggaggaaat ctgagtcgac ctgcagttcg
     2821 aagttcctat tctctagaaa gtataggaac ttcagagcgc ttttgaagct cacgctgccg
     2881 caagcactca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca
     2941 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
     3001 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
     3061 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
     3121 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     3181 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3241 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3301 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3361 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3421 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3481 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3541 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3601 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3661 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3721 agccgaactg ttcgccaggc tcaaggcgcg tatgcccgac ggcgaggatc tcgtcgtgac
     3781 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3841 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3901 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3961 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctaataagg
     4021 ggatcttgaa gttcctattc cgaagttcct attctctaga aagtatagga acttcgaagc
     4081 agctccagcc tacagcatgc cggtttgctg taaagtttag ggagatttga tggcttactc
     4141 tgttcaaaag tcgcgcctgg caaaggttgc gggtgtttcg cttgttttat tactcgctgc
     4201 ctgtagttct gactcacgct ataagcgtca ggtcagtggt gatgaagcct acctggaagc
     4261 ggcaccgctt gcggagcttc atgccccggc tggaatgatt ttgccggtga cctccggtga
     4321 ttatgcaatc ccggtgacca acggtagtgg tgctgtcggt aaggcgctgg acattcacta
     4381 gtcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     4441 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaggggtc cccaataatt
     4501 acgatttaaa ttagtagccc gcctaatgag cgggcttttt tttaattccc ctatttgttt
     4561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4621 tcaataatat tgaaaaagga agagtatgag cattcagcat tttcgtgtgg cgctgattcc
     4681 gttttttgcg gcgttttgcc tgccggtgtt tgcgcatccg gaaaccctgg tgaaagtgaa
     4741 agatgcggaa gatcaactgg gtgcgcgcgt gggctatatt gaactggatc tgaacagcgg
     4801 caaaattctg gaatcttttc gtccggaaga acgttttccg atgatgagca cctttaaagt
     4861 gctgctgtgc ggtgcggttc tgagccgtgt ggatgcgggc caggaacaac tgggccgtcg
     4921 tattcattat agccagaacg atctggtgga atatagcccg gtgaccgaaa aacatctgac
     4981 cgatggcatg accgtgcgtg aactgtgcag cgcggcgatt accatgagcg ataacaccgc
     5041 ggcgaacctg ctgctgacga ccattggcgg tccgaaagaa ctgaccgcgt ttctgcataa
     5101 catgggcgat catgtgaccc gtctggatcg ttgggaaccg gaactgaacg aagcgattcc
     5161 gaacgatgaa cgtgatacca ccatgccggc agcaatggcg accaccctgc gtaaactgct
     5221 gacgggtgag ctgctgaccc tggcaagccg ccagcaactg attgattgga tggaagcgga
     5281 taaagtggcg ggtccgctgc tgcgtagcgc gctgccggct ggctggttta ttgcggataa
     5341 aagcggtgcg ggcgaacgtg gcagccgtgg cattattgcg gcgctgggcc cggatggtaa
     5401 accgagccgt attgtggtga tttataccac cggcagccag gcgacgatgg atgaacgtaa
     5461 ccgtcagatt gcggaaattg gcgcgagcct gattaaacat tggtaaaccg atacaattaa
     5521 aggctccttt tggagccttt ttttttggac gacccttgtc cttttccgct gcataaccct
     5581 gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag
     5641 gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca
     5701 ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc
     5761 acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccgtaggcc ggccgcgatg
     5821 caggtggctg ctgaaccccc agccggaact gaccccacaa ggccctagcg tttgcaatgc
     5881 accaggtcat cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag
     5941 gcttcgccga cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc gcacatgagg
     6001 cggaaggttt ccagcttgag cgggtacggc tcccggtgcg agctgaaata gtcgaacatc
     6061 cgtcgggccg tcggcgacag cttgcggtac ttctcccata tgaatttcgt gtagtggtcg
     6121 ccagcaaaca gcacgacgat ttcctcgtcg atcaggacct ggcaacggga cgttttcttg
     6181 ccacggtcca ggacgcggaa gcggtgcagc agcgacaccg attccaggtg cccaacgcgg
     6241 tcggacgtga agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg
     6301 taataccggc cattgatcga ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg
     6361 atcggctcgc cgataggggt gcgcttcgcg tactccaaca cctgctgcca caccagttcg
     6421 tcatcgtcgg cccgcagctc gacgccggtg taggtgatct tcacgtcctt gttgacgtgg
     6481 aaaatgacct tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg
     6541 gcagagcggg ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg
     6601 aacaaggaaa gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc
     6661 ttggcttcgc tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc
     6721 atagttcctc gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc ctgttcgaga
     6781 cgacgcgaac gctccacggc ggccgatggc gcgggcaggg cagggggagc cagttgcacg
     6841 ctgtcgcgct cgatcttggc cgtagcttgc tggactatcg agccgacgga ctggaaggtt
     6901 tcgcggggcg cacgcatgac ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac
     6961 cccgcgtcga tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt cattcaccct
     7021 ccttgcggga ttgccccgga attaattccc cggatcgatc cgtcgatctt gatcccctgc
     7081 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     7141 taccagaggg cgccccagct ggcaattccg gttcgcttgc tgtccataaa accgcccagt
     7201 ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt
     7261 cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac
     7321 tggctttcta cgtggctgcc atttttgggg tgaggccgtt cgcggccgag gggcgcagcc
     7381 cctgggggga tgggaggccc gcgttagcgg gccgggaggg ttcgagaagg gggggcaccc
     7441 cccttcggcg tgcgcggtca cgcgcacagg gcgcagccct ggttaaaaac aaggtttata
     7501 aatattggtt taaaagcagg ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa
     7561 cccttgcaaa tgctggattt tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc
     7621 cctcatctgt cagcactctg cccctcaagt gtcaaggatc gcgcccctca tctgtcagta
     7681 gtcgcgcccc tcaagtgtca ataccgcagg gcacttatcc ccaggcttgt ccacatcatc
     7741 tgtgggaaac tcgcgtaaaa tcaggcgttt tcgccgattt gcgaggctgg ccagctccac
     7801 gtcgccggcc gaaatcgagc ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc
     7861 cctcaagtgt caacgtccgc ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat
     7921 ccacaacgcc ggcggcccta catggctctg ctgtagtgag tgggttgcgc tccggcagcg
     7981 gtcctgatcc cccgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacggcg
     8041 cgcccagctg tctagggcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     8101 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcct
//
