LOCUS       sequence-314741-        7059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A mutational reporter plasmid that can gain Kanamycin resistance 
            upon mutation. Contains two T7 promoters flanking the kanamycin 
            resistance gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7059)
  AUTHORS   Moore CL, Papa LJ 3rd, Shoulders MD
  TITLE     A Processive Protein Chimera Introduces Mutations across Defined DNA
            Regions In Vivo.
  JOURNAL   J Am Chem Soc. 2018 Sep 19;140(37):11560-11564. doi: 
            10.1021/jacs.8b04001. Epub 2018 Jul 18.
  PUBMED    29991261
REFERENCE   2  (bases 1 to 7059)
  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"; doi: 
            "10.1021/jacs.8b04001"; journalName: "J Am Chem Soc"; date: 
            "2018-09-19- 19"; volume: "140"; issue: "37"; pages: "11560-11564"
COMMENT     Sequence Label: Dual opposing T7 promoters reporter plasmid
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..27)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    308..781
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(854..1825)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(1825..3000)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    3326..3576
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(3579..4334)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      4425..4644
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             complement(join(4870..5661,5662..5730))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5493..5512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5731..5835)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5944..5962
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6006..6053
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6075..6875
                     /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="GSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6135..6154)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6745..6764
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(6954..6972)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
       61 ttagacgtca ggtggcactt ttcggggaaa tgtgcccggt taacgtgccg gcacggcctg
      121 ggtaaccagg tattttgtcc acataaccgt gcgcaaaatg ttgtggataa gcaggacaca
      181 gcagcaatcc acagcaggca tacaaccgca caccgaggtt actccgttct acaggttacg
      241 acgacatgtc aatacttgcc cttgacaggc attgatggaa tcgtagtctc acgctgatag
      301 tctgatcgac aatacaagtg ggaccgtggt cccagaccga taatcagacc gacaacacga
      361 gtgggatcgt ggtcccagac taataatcag accgacgata cgagtgggac cgtggtccca
      421 gactaataat cagaccgacg atacgagtgg gaccgtggtt ccagactaat aatcagaccg
      481 acgatacgag tgggaccgtg gtcccagact aataatcaga ccgacgatac gagtgggacc
      541 atggtcccag actaataatc agaccgacga tacgagtggg accgtggtcc cagtctgatt
      601 atcagaccga cgatacgagt gggaccgtgg tcccagacta ataatcagac cgacgatacg
      661 agtgggaccg tggtcccaga ctaataatca gaccgacgat acgagtggga ccgtggtccc
      721 agtctgatta tcagaccgac gatacaagtg gaacagtggg cccagagaga atattcaggc
      781 cagttatgct ttctggcctg taacaaagga cattaagtaa agacagataa acgtagacta
      841 aaacgtggtc gcatcagggt gctggctttt caagttcctt aagaatggcc tcaattttct
      901 ctatacactc agttggaaca cgagacctgt ccaggttaag caccatttta tcgcccttat
      961 acaatactgt cgctccagga gcaaactgat gtcgtgagct taaactagtt cttgatgcag
     1021 atgacgtttt aagcacagaa gttaaaagag tgataacttc ttcagcttca aatatcaccc
     1081 cagctttttt ctgctcatga aggttagatg cctgctgctt aagtaattcc tctttatctg
     1141 taaaggcttt ttgaagtgca tcacctgacc gggcagatag ttcaccgggg tgagaaaaaa
     1201 gagcaacaac tgatttaggc aatttggcgg tgttgataca gcgggtaata atcttacgtg
     1261 aaatattttc cgcatcagcc agcgcagaaa tatttccagc aaattcattc tgcaatcggc
     1321 ttgcataacg ctgaccacgt tcataagcac ttgttgggcg ataatcgtta cccaatctgg
     1381 ataatgcagc catctgctca tcatccagct cgccaaccag aacacgataa tcactttcgg
     1441 taagtgcagc agctttacga cggcgactcc catcggcaat ttctatgaca ccagatactc
     1501 ttcgaccgaa cgccggtgtc tgttgaccag tcagtagaaa agaagggatg agatcatcca
     1561 gtgcgtcctc agtaagcagc tcctggtcac gttcattacc tgaccatacc cgagaggtct
     1621 tctcaacact atcaccccgg agcacttcaa gagtaaactt cacatcccga ccacatacag
     1681 gcaaagtaat ggcattaccg cgagccatta ctcctacgcg cgcaattaac gaatccacca
     1741 tcggggcagc tggtgtcgat aacgaagtat cttcaaccgg ttgagtattg agcgtatgtt
     1801 ttggaataac aggcgcacgc ttcattatct aatctcccag cgtggtttaa tcagacgatc
     1861 gaaaatttca ttgcagacag gttcccaaat agaaagagca tttctccagg caccagttga
     1921 agagcgttga tcaatggcct gttcaaaaac agttctcatc cggatctgac ctttaccaac
     1981 ttcatccgtt tcacgtacaa cattttttag aaccatgctt ccccaggcat cccgaatttg
     2041 ctcctccatc cacggggact gagagccatt actattgctg tatttggtaa gcaaaatacg
     2101 tacatcaggc tcgaaccctt taagatcaac gttcttgagc agatcacgaa gcatatcgaa
     2161 aaactgcagt gcggaggtgt agtcaaacaa ctcagcaggc gtgggaacaa tcagcacatc
     2221 agcagcacat acgacattaa tcgtgccgat acccaggtta ggcgcgctgt caataactat
     2281 gacatcatag tcatgagcaa cagtttcaat ggccagtcgg agcatcaggt gtggatcggt
     2341 gggcagttta ccttcatcaa atttgcccat taactcagtt tcaatacggt gcagagccag
     2401 acaggaagga ataatgtcaa gccccggcca gcaagtgggc tttattgcat aagtgacatc
     2461 gtccttttcc ccaagataga aaggcaggag agtgtcttct gcatgaatat gaagatctgg
     2521 tacccatccg tgatacattg aggctgttcc ctgggggtcg ttaccttcca cgagcaaaac
     2581 acgtagcccc ttcagagcca gatcctgagc aagatgaaca gaaactgagg ttttgtaaac
     2641 gccaccttta tgggcagcaa ccccgatcac cggtggaaat acgtcttcag cacgtcgcaa
     2701 tcgcgtacca aacacatcac gcatatgatt aatttgttca attgtataac caacacgttg
     2761 ctcaacccgt cctcgaattt ccatatccgg gtgcggtagt cgccctgctt tctcggcatc
     2821 tctgatagcc tgagaagaaa ccccaactaa atccgctgct tcacctattc tccagcgccg
     2881 ggttattttc ctcgcttccg ggctgtcatc attaaactgt gcaatggcga tagccttcgt
     2941 catttcatga ccagcgttta tgcactggtt aagtgtttcc atgagtttca ttctgaacat
     3001 cctttaatca ttgctttgcg tttttttatt aaatcttgca atttactgca aagcaacaac
     3061 aaaatcgcaa agtcatcaaa aaaccgcaaa gttgtttaaa ataagagcaa cactacaaaa
     3121 ggagataaga agagcacata cctcagtcac ttattatcac tagcgctcgc cgcagccgtg
     3181 taaccgagca tagcgagcga actggcgagg aagcaaagaa gaactgttct gtcagatagc
     3241 tcttacgctc agcgcaagaa gaaatatcca ccgtgggaaa aactccaggt agaggtacac
     3301 acgcggatag ccaattcaga gtaataaact gtgataatca accctcatca atgatgacga
     3361 actaaccccc gatatcaggt cacatgacga agggaaagag aaggaaatca actgtgacaa
     3421 actgccctca aatttggctt ccttaaaaat tacagttcaa aaagtatgag aaaatccatg
     3481 caggctgaag gaaacagcaa aactgtgaca aattaccctc agtaggtcag aacaaatgtg
     3541 acgaaccacc ctcaaatctg tgacagataa ccctcagact atcctgtcgt catggaagtg
     3601 atatcgcgga aggaaaatac gatatgagtc gtctggcggc ctttcttttt ctcaatgtat
     3661 gagaggcgca ttggagttct gctgttgatc tcattaacac agacctgcag gaagcggcgg
     3721 cggaagtcag gcatacgctg gtaactttga ggcagctggt aacgctctat gatccagtcg
     3781 attttcagag agacgatgcc tgagccatcc ggcttacgat actgacacag ggattcgtat
     3841 aaacgcatgg catacggatt ggtgatttct tttgtttcac taagccgaaa ctgcgtaaac
     3901 cggttctgta acccgataaa gaagggaatg agatatgggt tgatatgtac actgtaaagc
     3961 cctctggatg gactgtgcgc acgtttgata aaccaaggaa aagattcata gcctttttca
     4021 tcgccggcat cctcttcagg gcgataaaaa accacttcct tccccgcgaa actcttcaat
     4081 gcctgccgta tatccttact ggcttccgca gaggtcaatc cgaatatttc agcatattta
     4141 gcaacatgga tctcgcagat accgtcatgt tcctgtaggg tgccatcaga ttttctgatc
     4201 tggtcaacga acagatacag catacgtttt tgatcccggg agagactata tgccgcctca
     4261 gtgaggtcgt ttgactggac gattcgcggg ctatttttac gtttcttgtg attgataacc
     4321 gctgtttccg ccatgacaga tccatgtgaa gtgtgacaag tttttagatt gtcacactaa
     4381 ataaaaaaga gtcaataagc agggataact ttgtgaaaaa acagcttctt ctgagggcaa
     4441 tttgtcacag ggttaagggc aatttgtcac agacaggact gtcatttgag ggtgatttgt
     4501 cacactgaaa gggcaatttg tcacaacacc ttctctagaa ccagcatgga taaaggccta
     4561 caaggcgctc taaaaaagaa gatctaaaaa ctataaaaaa aataattata aaaatatccc
     4621 cgtggataag tggataaccc caagggaagt tttttcaggc atcgtgtgta agcagaatat
     4681 ataagtgctg ttccctggtg cttcctcgct cactcgagct acggggtctg acgctcagtg
     4741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct ggcggccgcc tttcagcaaa
     5881 aaaccccgcg agacccccga agaggccccg cggggttatg ctaggtcgac ggagctcgaa
     5941 ttctaatacg actcactata gggagaccca agctggcttg acaattaatc atcggctcga
     6001 agcttgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     6061 ggaactaaac cacgggatcg gccattgaac aagatggatt gcacgcaggt tctccggccg
     6121 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     6181 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     6241 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     6301 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     6361 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6421 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6481 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6541 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6601 tcaaggcgcg catgcccgac ggcgatgatc tcgtcgtgac ccatggcgat gcctgcttgc
     6661 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6721 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6781 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6841 tcgccttcta tcgccttctt gacgagttct tctgagggga tcaattctct agagcttaat
     6901 taacgcagcc tgaatggcga atagaagttt aaacgctagc cagcttgggt ctccctatag
     6961 tgagtcgtat tatcgagctc cgtcgaccta gcataacccc gcggggcctc ttcgggggtc
     7021 tcgcggggtt ttttgctgaa agggatccaa ttcttgaag
//
