LOCUS       sequence-311166-        8339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A mutational reporter plasmid that can gain Streptomycin and/or 
            Tetracycline resistance upon mutation. Contains T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8339)
  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 8339)
  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: T7 promoter + rpsL reporter plasmid
FEATURES             Location/Qualifiers
     source          1..8339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    252..725
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(798..1769)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(1769..2944)
                     /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    3270..3520
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(3523..4278)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      4369..4588
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             complement(join(4814..5605,5606..5674))
                     /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     5437..5456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5675..5779)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5888..5907
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5888..5906
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        join(5913..5918,5919..5934,5935..5941)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        join(6990..6995,6996..7012,7013..7018)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             7064..8266
                     /codon_start=1
                     /gene="tetA from transposon Tn10"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="NSSTKIALVITLLDAMGIGLIMPVLPTLLREFIASEDIANHFGVL
                     LALYALMQVIFAPWLGKMSDRFGRRPVLLLSLIGASLDYLLLAFSSALWMLYLGRLLSG
                     ITGATGAVAASVIADTTSASQRVKWFGWLGASFGLGLIAGPIIGGFAGEISPHSPFFIA
                     ALLNIVTFLVVMFWFRETKNTRDNTDTEVGVETQSNSVYITLFKTMPILLIIYFSAQLI
                     GQIPATVWVLFTENRFGWNSMMVGFSLAGLGLLHSVFQAFVAGRIATKWGEKTAVLLGF
                     IADSSAFAFLAFISEGWLVFPVLILLAGGGIALPALQGVMSIQTKSHQQGALQGLLVSL
                     TNATGVIGPLLFAVIYNHSLPIWDGWIWIIGLAFYCIIILLSMTFMLTPQAQGSKQETS
                     A"
     primer_bind     complement(8292..8310)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttcttagac gtcaggtggc acttttcggg gaaatgtgcc cggttaacgt gccggcacgg
       61 cctgggtaac caggtatttt gtccacataa ccgtgcgcaa aatgttgtgg ataagcagga
      121 cacagcagca atccacagca ggcatacaac cgcacaccga ggttactccg ttctacaggt
      181 tacgacgaca tgtcaatact tgcccttgac aggcattgat ggaatcgtag tctcacgctg
      241 atagtctgat cgacaataca agtgggaccg tggtcccaga ccgataatca gaccgacaac
      301 acgagtggga tcgtggtccc agactaataa tcagaccgac gatacgagtg ggaccgtggt
      361 cccagactaa taatcagacc gacgatacga gtgggaccgt ggttccagac taataatcag
      421 accgacgata cgagtgggac cgtggtccca gactaataat cagaccgacg atacgagtgg
      481 gaccatggtc ccagactaat aatcagaccg acgatacgag tgggaccgtg gtcccagtct
      541 gattatcaga ccgacgatac gagtgggacc gtggtcccag actaataatc agaccgacga
      601 tacgagtggg accgtggtcc cagactaata atcagaccga cgatacgagt gggaccgtgg
      661 tcccagtctg attatcagac cgacgataca agtggaacag tgggcccaga gagaatattc
      721 aggccagtta tgctttctgg cctgtaacaa aggacattaa gtaaagacag ataaacgtag
      781 actaaaacgt ggtcgcatca gggtgctggc ttttcaagtt ccttaagaat ggcctcaatt
      841 ttctctatac actcagttgg aacacgagac ctgtccaggt taagcaccat tttatcgccc
      901 ttatacaata ctgtcgctcc aggagcaaac tgatgtcgtg agcttaaact agttcttgat
      961 gcagatgacg ttttaagcac agaagttaaa agagtgataa cttcttcagc ttcaaatatc
     1021 accccagctt ttttctgctc atgaaggtta gatgcctgct gcttaagtaa ttcctcttta
     1081 tctgtaaagg ctttttgaag tgcatcacct gaccgggcag atagttcacc ggggtgagaa
     1141 aaaagagcaa caactgattt aggcaatttg gcggtgttga tacagcgggt aataatctta
     1201 cgtgaaatat tttccgcatc agccagcgca gaaatatttc cagcaaattc attctgcaat
     1261 cggcttgcat aacgctgacc acgttcataa gcacttgttg ggcgataatc gttacccaat
     1321 ctggataatg cagccatctg ctcatcatcc agctcgccaa ccagaacacg ataatcactt
     1381 tcggtaagtg cagcagcttt acgacggcga ctcccatcgg caatttctat gacaccagat
     1441 actcttcgac cgaacgccgg tgtctgttga ccagtcagta gaaaagaagg gatgagatca
     1501 tccagtgcgt cctcagtaag cagctcctgg tcacgttcat tacctgacca tacccgagag
     1561 gtcttctcaa cactatcacc ccggagcact tcaagagtaa acttcacatc ccgaccacat
     1621 acaggcaaag taatggcatt accgcgagcc attactccta cgcgcgcaat taacgaatcc
     1681 accatcgggg cagctggtgt cgataacgaa gtatcttcaa ccggttgagt attgagcgta
     1741 tgttttggaa taacaggcgc acgcttcatt atctaatctc ccagcgtggt ttaatcagac
     1801 gatcgaaaat ttcattgcag acaggttccc aaatagaaag agcatttctc caggcaccag
     1861 ttgaagagcg ttgatcaatg gcctgttcaa aaacagttct catccggatc tgacctttac
     1921 caacttcatc cgtttcacgt acaacatttt ttagaaccat gcttccccag gcatcccgaa
     1981 tttgctcctc catccacggg gactgagagc cattactatt gctgtatttg gtaagcaaaa
     2041 tacgtacatc aggctcgaac cctttaagat caacgttctt gagcagatca cgaagcatat
     2101 cgaaaaactg cagtgcggag gtgtagtcaa acaactcagc aggcgtggga acaatcagca
     2161 catcagcagc acatacgaca ttaatcgtgc cgatacccag gttaggcgcg ctgtcaataa
     2221 ctatgacatc atagtcatga gcaacagttt caatggccag tcggagcatc aggtgtggat
     2281 cggtgggcag tttaccttca tcaaatttgc ccattaactc agtttcaata cggtgcagag
     2341 ccagacagga aggaataatg tcaagccccg gccagcaagt gggctttatt gcataagtga
     2401 catcgtcctt ttccccaaga tagaaaggca ggagagtgtc ttctgcatga atatgaagat
     2461 ctggtaccca tccgtgatac attgaggctg ttccctgggg gtcgttacct tccacgagca
     2521 aaacacgtag ccccttcaga gccagatcct gagcaagatg aacagaaact gaggttttgt
     2581 aaacgccacc tttatgggca gcaaccccga tcaccggtgg aaatacgtct tcagcacgtc
     2641 gcaatcgcgt accaaacaca tcacgcatat gattaatttg ttcaattgta taaccaacac
     2701 gttgctcaac ccgtcctcga atttccatat ccgggtgcgg tagtcgccct gctttctcgg
     2761 catctctgat agcctgagaa gaaaccccaa ctaaatccgc tgcttcacct attctccagc
     2821 gccgggttat tttcctcgct tccgggctgt catcattaaa ctgtgcaatg gcgatagcct
     2881 tcgtcatttc atgaccagcg tttatgcact ggttaagtgt ttccatgagt ttcattctga
     2941 acatccttta atcattgctt tgcgtttttt tattaaatct tgcaatttac tgcaaagcaa
     3001 caacaaaatc gcaaagtcat caaaaaaccg caaagttgtt taaaataaga gcaacactac
     3061 aaaaggagat aagaagagca catacctcag tcacttatta tcactagcgc tcgccgcagc
     3121 cgtgtaaccg agcatagcga gcgaactggc gaggaagcaa agaagaactg ttctgtcaga
     3181 tagctcttac gctcagcgca agaagaaata tccaccgtgg gaaaaactcc aggtagaggt
     3241 acacacgcgg atagccaatt cagagtaata aactgtgata atcaaccctc atcaatgatg
     3301 acgaactaac ccccgatatc aggtcacatg acgaagggaa agagaaggaa atcaactgtg
     3361 acaaactgcc ctcaaatttg gcttccttaa aaattacagt tcaaaaagta tgagaaaatc
     3421 catgcaggct gaaggaaaca gcaaaactgt gacaaattac cctcagtagg tcagaacaaa
     3481 tgtgacgaac caccctcaaa tctgtgacag ataaccctca gactatcctg tcgtcatgga
     3541 agtgatatcg cggaaggaaa atacgatatg agtcgtctgg cggcctttct ttttctcaat
     3601 gtatgagagg cgcattggag ttctgctgtt gatctcatta acacagacct gcaggaagcg
     3661 gcggcggaag tcaggcatac gctggtaact ttgaggcagc tggtaacgct ctatgatcca
     3721 gtcgattttc agagagacga tgcctgagcc atccggctta cgatactgac acagggattc
     3781 gtataaacgc atggcatacg gattggtgat ttcttttgtt tcactaagcc gaaactgcgt
     3841 aaaccggttc tgtaacccga taaagaaggg aatgagatat gggttgatat gtacactgta
     3901 aagccctctg gatggactgt gcgcacgttt gataaaccaa ggaaaagatt catagccttt
     3961 ttcatcgccg gcatcctctt cagggcgata aaaaaccact tccttccccg cgaaactctt
     4021 caatgcctgc cgtatatcct tactggcttc cgcagaggtc aatccgaata tttcagcata
     4081 tttagcaaca tggatctcgc agataccgtc atgttcctgt agggtgccat cagattttct
     4141 gatctggtca acgaacagat acagcatacg tttttgatcc cgggagagac tatatgccgc
     4201 ctcagtgagg tcgtttgact ggacgattcg cgggctattt ttacgtttct tgtgattgat
     4261 aaccgctgtt tccgccatga cagatccatg tgaagtgtga caagttttta gattgtcaca
     4321 ctaaataaaa aagagtcaat aagcagggat aactttgtga aaaaacagct tcttctgagg
     4381 gcaatttgtc acagggttaa gggcaatttg tcacagacag gactgtcatt tgagggtgat
     4441 ttgtcacact gaaagggcaa tttgtcacaa caccttctct agaaccagca tggataaagg
     4501 cctacaaggc gctctaaaaa agaagatcta aaaactataa aaaaaataat tataaaaata
     4561 tccccgtgga taagtggata accccaaggg aagttttttc aggcatcgtg tgtaagcaga
     4621 atatataagt gctgttccct ggtgcttcct cgctcactcg agctacgggg tctgacgctc
     4681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4801 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4861 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4921 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4981 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5041 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5101 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5161 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5221 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5281 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5341 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5401 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5461 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5521 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5581 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5641 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5701 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5761 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctggcggc cgcctttcag
     5821 caaaaaaccc cgcgagaccc ccgaagaggc cccgcggggt tatgctaggt cgacggagct
     5881 cgaattctaa tacgactcac tatagggaga ccttgacaat taatcatcgg ctcgtataat
     5941 gccagaagct tagcaaaagc taaaaccagg agctatttaa tggcaacagt taaccagctg
     6001 gtacgcaaac cacgtgctcg caaagttgcg aaaagcaacg tgcctgcgct ggaagcatgc
     6061 ccgcaaaaac gtggcgtatg tactcgtgta tatactacca ctcctaaaaa accgaactcc
     6121 gcgctgcgta aagtatgccg tgttcgtctg actaacggtt tcgaagtgac ttcctacatc
     6181 ggtggtgaag gtcacaacct gcaggagcac tccgtgatcc tgatccgtgg cggtcgtgtt
     6241 aaagacctcc cgggtgttcg ttaccacacc gtacgtggtg cgcttgactg ctccggcgtt
     6301 aaagaccgta agcaggctcg ttccaagtat ggcgtgaagc gtcctaaggc ttaatggttt
     6361 aattaacgca gcctgaatgg cgaatagaag tttaaacgct agacctagca taaccccgcg
     6421 gggcctcttc gggggtctcg cggggttttt tgctgaaagg ctagacctag cataaccccg
     6481 cggggcctct tcgggggtct cgcggggttt tttgctgaaa ggctagacct agcataaccc
     6541 cgcggggcct cttcgggggt ctcgcggggt tttttgctga aaggctagac ctagcataac
     6601 cccgcggggc ctcttcgggg gtctcgcggg gttttttgct gaaaggctag acctagcata
     6661 accccgcggg gcctcttcgg gggtctcgcg gggttttttg ctgaaaggct aggaagttta
     6721 aacgctagac ctagcataac cccgcggggc ctcttcgggg gtctcgcggg gttttttgct
     6781 gaaaggctag acctagcata accccgcggg gcctcttcgg gggtctcgcg gggttttttg
     6841 ctgaaaggct agacctagca taaccccgcg gggcctcttc gggggtctcg cggggttttt
     6901 tgctgaaagg ctagacctag cataaccccg cggggcctct tcgggggtct cgcggggttt
     6961 tttgctgaaa ggctaggata tattgatcct tgacagctta tcatcgataa gctttaatgc
     7021 ggtagtttat cacagttgct aacgcagtca ggcaccgtgt acgaatagtt cgacaaagat
     7081 cgcattggta attacgttac tcgatgccat ggggattggc cttatcatgc cagtcttgcc
     7141 aacgttatta cgtgaattta ttgcttcgga agatatcgct aaccactttg gcgtattgct
     7201 tgcactttat gcgttaatgc aggttatctt tgctccttgg cttggaaaaa tgtctgaccg
     7261 atttggtcgg cgcccagtgc tgttgttgtc attaataggc gcatcgctgg attacttatt
     7321 gctggctttt tcaagtgcgc tttggatgct gtatttaggc cgtttgcttt cagggatcac
     7381 aggagctact ggggctgtcg cggcatcggt cattgccgat accacctcag cttctcaacg
     7441 cgtgaagtgg ttcggttggt taggggcaag ttttgggctt ggtttaatag cggggcctat
     7501 tattggtggt tttgcaggag agatttcacc gcatagtccc ttttttatcg ctgcgttgct
     7561 aaatattgtc actttccttg tggttatgtt ttggttccgt gaaaccaaaa atacacgtga
     7621 taatacagat accgaagtag gggttgagac gcaatcgaat tcggtataca tcactttatt
     7681 taaaacgatg cccattttgt tgattattta tttttcagcg caattgatag gccaaattcc
     7741 cgcaacggtg tgggtgctat ttaccgaaaa tcgttttgga tggaatagca tgatggttgg
     7801 cttttcatta gcgggtcttg gtcttttaca ctcagtattc caagcctttg tggcaggaag
     7861 aatagccact aaatggggcg aaaaaacggc agtactgctc ggatttattg cagatagtag
     7921 tgcatttgcc tttttagcgt ttatatctga aggttggtta gttttccctg ttttaatttt
     7981 attggctggt ggtgggatcg ctttacctgc attacaggga gtgatgtcta tccaaacaaa
     8041 gagtcatcag caaggtgctt tacagggatt attggtgagc cttaccaatg caaccggtgt
     8101 tattggccca ttactgtttg ctgttattta taatcattca ctaccaattt gggatggctg
     8161 gatttggatt attggtttag cgttttactg tattattatc ctgctatcga tgaccttcat
     8221 gttaacccct caagctcagg ggagtaaaca ggagacaagt gcttagtgat ccaattcttg
     8281 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatgg
//
