LOCUS       sequence-203820-        5362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse monomeric Rev7 protein with mutations abolishing 
            binding to Champ1 and Rev3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5362)
  AUTHORS   Thanos Halazonetis
  TITLE     Expression plasmids for production of Rev7 and Champ1 proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5362)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pT5T_Rev7_1-211_SA112_KVK_YAK
FEATURES             Location/Qualifiers
     source          1..5362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..64
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        45..63
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             95..117
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             125..157
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     primer_bind     complement(895..913)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      909..956
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1001..1019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1087..1191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1192..1260,1261..2052)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRXDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1410..1429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2223..2810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2711..2730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2964..2981
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2996..3135
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3061..3080)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3221..3243
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3237..3428)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4067..5257)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     4886..4905
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(5305..5310,5311..5327,5328..5333))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 tagcgaaagc taaggatttt ttttagatct cgatcccgcg aaattaatac gactcactat
       61 agggagacca caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat
      121 acatatggct agcatgactg gtggacagca aatgggtcgg gatccattgg aggatgatta
      181 aatgaccacc ctcacgcgcc aagacctcaa ctctgcccaa gtggtggctg acgtgctctc
      241 cgagttcctg gaggtggccg tgcacctgat tctctatgtg cgcgaggtct acccggtggg
      301 ctacttccag aagcgcaaga agtacaacgt gccggttcag atgtcctgtc acccggagct
      361 gaaccagtac atccaggaca cactccactg cgtcaaacct ctcctggaga agaacgatgt
      421 ggagaaggtg gtggtggtga ttttggataa ggaacaccgc ccagtggaga agtttgtctt
      481 tgagatcact cagcctccct tgctgtccat caattcagac tccctcctgt ctcatgtgga
      541 gcagctgctt cgagccttca tccttaagat tagtaaggtt gataaggtcc tggatcacaa
      601 ccctccaggc tgcacattta cagtcctcgt gcacacaaga gaagctgcta ctcgaaacgc
      661 ggagaagaag caggtcatca aggacttccc gtggattctg gcagatgaac aggatgtcca
      721 catgcacgac ccccgcttga tacccctaaa aaccatgacg tcggacattt taaagatgca
      781 gctctacgtt gaagagcgag cgcataagaa cagctaagcg gccgcagctt ggtaccgact
      841 agtaagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      901 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      961 aaggaggaac tatatccgga taattcttga agacgaaagg gcctcgtgat acgcctattt
     1021 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1081 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1141 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1201 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1261 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1321 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1381 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtrttgac
     1441 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1501 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1561 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1621 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     1681 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     1741 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     1801 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     1861 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     1921 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1981 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2041 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa ccaccgctac
     2281 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2341 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2401 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2461 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2521 aggagcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2581 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2641 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2701 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2761 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2821 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2881 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2941 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3001 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3061 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3121 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3181 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3241 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3301 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     3361 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     3421 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     3481 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     3541 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     3601 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     3661 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     3721 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     3781 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     3841 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     3901 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     3961 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     4021 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     4081 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     4141 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
     4201 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
     4261 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
     4321 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
     4381 agacgtagcc cagcgcgtcg gcagccatgc cggcgataat ggcctgcttc tcgccgaaac
     4441 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     4501 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     4561 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     4621 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4681 gcatcggtcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     4741 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     4801 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     4861 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     4921 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gagaatccac aggacgggtg
     4981 tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc
     5041 ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg
     5101 catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc
     5161 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg
     5221 tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc
     5281 aatttaactg tgataaacta ccgcattaaa acttatcgat gataagctgt caaacatgag
     5341 aattgagctc cgcggagatc ct
//
