LOCUS       sequence-203916-        5362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse monomeric Rev7 protein.
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  JOURNAL   Unpublished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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..5362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..68
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        49..67
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             99..121
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             129..161
                     /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(899..917)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      913..960
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1005..1023)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1091..1195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1196..1264,1265..2056)
                     /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(1414..1433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2227..2814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2715..2734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2968..2985
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3000..3139
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3065..3084)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3225..3247
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3241..3432)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4071..5261)
                     /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     4890..4909
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(5309..5314,5315..5331,5332..5337))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 tccttagcga aagctaagga ttttttttag atctcgatcc cgcgaaatta atacgactca
       61 ctatagggag accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag
      121 atatacatat ggctagcatg actggtggac agcaaatggg tcgggatcca ttggaggatg
      181 attaaatgac caccctcacg cgccaagacc tcaactctgc ccaagtggtg gctgacgtgc
      241 tctccgagtt cctggaggtg gccgtgcacc tgattctcta tgtgcgcgag gtctacccgg
      301 tgggcatctt ccagaagcgc aagaagtaca acgtgccggt tcagatgtcc tgtcacccgg
      361 agctgaacca gtacatccag gacacactcc actgcgtcaa acctctcctg gagaagaacg
      421 atgtggagaa ggtggtggtg gtgattttgg ataaggaaca ccgcccagtg gagaagtttg
      481 tctttgagat cactcagcct cccttgctgt ccatcaattc agactccctc ctgtctcatg
      541 tggagcagct gcttcgagcc ttcatcctta agattagtaa ggttgataag gtcctggatc
      601 acaaccctcc aggctgcaca tttacagtcc tcgtgcacac aagagaagct gctactcgaa
      661 acatggagaa gatacaggtc atcaaggact tcccgtggat tctggcagat gaacaggatg
      721 tccacatgca cgacccccgc ttgatacccc taaaaaccat gacgtcggac attttaaaga
      781 tgcagctcta cgttgaagag cgagcgcata agaacagcta agcggccgca gcttggtacc
      841 gactagtaag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
      901 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      961 ctgaaaggag gaactatatc cggataattc ttgaagacga aagggcctcg tgatacgcct
     1021 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1081 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1381 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1441 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1501 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     1741 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1801 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2041 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaccaccg
     2281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2341 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2581 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2881 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2941 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3001 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
     3061 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     3121 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3181 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3241 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     3301 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     3361 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     3421 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     3481 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     3541 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     3601 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     3661 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     3721 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     3781 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     3841 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     3901 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat
     3961 ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc
     4021 tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag
     4081 gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg
     4141 cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg
     4201 atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt
     4261 ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg
     4321 ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc
     4381 agcaagacgt agcccagcgc gtcggcagcc atgccggcga taatggcctg cttctcgccg
     4441 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     4501 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     4561 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     4621 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     4681 aagggcatcg gtcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     4741 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     4801 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     4861 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     4921 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagagaat ccacaggacg
     4981 ggtgtggtcg ccatgatcgc gtagtcgata gtggctccaa gtagcgaagc gagcaggact
     5041 gggcggcggc caaagcggtc ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc
     5101 aacgcatata gcgctagcag cacgccatag tgactggcga tgctgtcgga atggacgata
     5161 tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg
     5221 acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt
     5281 tagcaattta actgtgataa actaccgcat taaaacttat cgatgataag ctgtcaaaca
     5341 tgagaattga gctccgcgga ga
//
