LOCUS       sequence-203205-        5497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse monomeric Rev7 and fragment of Rev3 protein (amino 
            acids 1873-1898) from bicistronic plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5497)
  AUTHORS   Thanos Halazonetis
  TITLE     Expression plasmids for production of Rev7 and Champ1 proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5497)
  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            "Unpublished"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_Rev3_1873-1898
FEATURES             Location/Qualifiers
     source          1..5497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..62
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        43..61
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             93..115
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             123..155
                     /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"
     RBS             835..857
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1028..1046)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1042..1089
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1134..1152)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1220..1324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1325..1393,1394..2185)
                     /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"
     primer_bind     complement(1543..1562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2356..2943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2844..2863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3097..3114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3129..3268
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3194..3213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3354..3376
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3370..3561)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4200..5390)
                     /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     5019..5038
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(5438..5443,5444..5460,5461..5466))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 gcgaaagcta aggatttttt ttagatctcg atcccgcgaa attaatacga ctcactatag
       61 ggagaccaca acggtttccc tctagaaata attttgttta actttaagaa ggagatatac
      121 atatggctag catgactggt ggacagcaaa tgggtcggga tccattggag gatgattaaa
      181 tgaccaccct cacgcgccaa gacctcaact ctgcccaagt ggtggctgac gtgctctccg
      241 agttcctgga ggtggccgtg cacctgattc tctatgtgcg cgaggtctac ccggtgggca
      301 tcttccagaa gcgcaagaag tacaacgtgc cggttcagat gtcctgtcac ccggagctga
      361 accagtacat ccaggacaca ctccactgcg tcaaacctct cctggagaag aacgatgtgg
      421 agaaggtggt ggtggtgatt ttggataagg aacaccgccc agtggagaag tttgtctttg
      481 agatcactca gcctcccttg ctgtccatca attcagactc cctcctgtct catgtggagc
      541 agctgcttcg agccttcatc cttaagatta gtaaggttga taaggtcctg gatcacaacc
      601 ctccaggctg cacatttaca gtcctcgtgc acacaagaga agctgctact cgaaacatgg
      661 agaagataca ggtcatcaag gacttcccgt ggattctggc agatgaacag gatgtccaca
      721 tgcacgaccc ccgcttgata cccctaaaaa ccatgacgtc ggacatttta aagatgcagc
      781 tctacgttga agagcgagcg cataagaaca gctaacccgg gctctagaaa taattttgtt
      841 taactttaag aaggagatat accatgggtc gtactgctaa catcctgaaa ccgcttatgt
      901 ctccaccgag tcgtgaagaa atcatggcta ctctgctgga tcatgactaa gcggccgcag
      961 cttggtaccg actagtaaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1021 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1081 ggttttttgc tgaaaggagg aactatatcc ggataattct tgaagacgaa agggcctcgt
     1141 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     1201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1321 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1381 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1441 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1501 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1561 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1621 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1681 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1741 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1801 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1861 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1921 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1981 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2041 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2101 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2161 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2221 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2281 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2401 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2461 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2521 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2581 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2641 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2701 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2761 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2821 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2881 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2941 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3001 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3061 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3121 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     3181 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3241 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     3301 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3361 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3421 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     3481 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     3541 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     3601 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     3661 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     3721 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     3781 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     3841 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     3901 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     3961 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     4021 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     4081 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     4141 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     4201 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     4261 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     4321 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     4381 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     4441 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     4501 gcgaacgcca gcaagacgta gcccagcgcg tcggcagcca tgccggcgat aatggcctgc
     4561 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     4621 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     4681 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     4741 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     4801 aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat taggaagcag
     4861 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     4921 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     4981 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     5041 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagagaatc
     5101 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
     5161 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
     5221 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
     5281 tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
     5341 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
     5401 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaaacttatc gatgataagc
     5461 tgtcaaacat gagaattgag ctccgcggag atcctta
//
