LOCUS       sequence-203917-        5501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse monomeric Rev7 and fragment of Champ1 protein (amino
            acids 328-355) from bicistronic plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5501)
  AUTHORS   Thanos Halazonetis
  TITLE     Expression plasmids for production of Rev7 and Champ1 proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5501)
  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            "Unpublished"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_Champ1_328-355
FEATURES             Location/Qualifiers
     source          1..5501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..76
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        57..75
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             107..129
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             137..169
                     /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             849..871
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1045..1063)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1059..1106
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1151..1169)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1237..1341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1342..1410,1411..2202)
                     /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(1560..1579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2373..2961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2862..2881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3115..3132
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3147..3286
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3212..3231)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3372..3394
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3388..3579)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4218..5408)
                     /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     5037..5056
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(5456..5461,5462..5478,5479..5484))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 cgcggagatc cttagcgaaa gctaaggatt ttttttagat ctcgatcccg cgaaattaat
       61 acgactcact atagggagac cacaacggtt tccctctaga aataattttg tttaacttta
      121 agaaggagat atacatatgg ctagcatgac tggtggacag caaatgggtc gggatccatt
      181 ggaggatgat taaatgacca ccctcacgcg ccaagacctc aactctgccc aagtggtggc
      241 tgacgtgctc tccgagttcc tggaggtggc cgtgcacctg attctctatg tgcgcgaggt
      301 ctacccggtg ggcatcttcc agaagcgcaa gaagtacaac gtgccggttc agatgtcctg
      361 tcacccggag ctgaaccagt acatccagga cacactccac tgcgtcaaac ctctcctgga
      421 gaagaacgat gtggagaagg tggtggtggt gattttggat aaggaacacc gcccagtgga
      481 gaagtttgtc tttgagatca ctcagcctcc cttgctgtcc atcaattcag actccctcct
      541 gtctcatgtg gagcagctgc ttcgagcctt catccttaag attagtaagg ttgataaggt
      601 cctggatcac aaccctccag gctgcacatt tacagtcctc gtgcacacaa gagaagctgc
      661 tactcgaaac atggagaaga tacaggtcat caaggacttc ccgtggattc tggcagatga
      721 acaggatgtc cacatgcacg acccccgctt gataccccta aaaaccatga cgtcggacat
      781 tttaaagatg cagctctacg ttgaagagcg agcgcataag aacagctaac ccgggctcta
      841 gaaataattt tgtttaactt taagaaggag atataccatg tcagcatcat caggaccttg
      901 gaagccagct aaacctgctc catctgtgtc tcctggacct tggaaaccaa ttccttctgt
      961 atcttaagcg gccgcagctt ggtaccgact agtaagatcc ggctgctaac aaagcccgaa
     1021 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     1081 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga taattcttga
     1141 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1201 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1261 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1321 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1381 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1441 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1501 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1561 ctatgtggcg cggtattatc ccgtrttgac gccgggcaag agcaactcgg tcgccgcata
     1621 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1681 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1741 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1801 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1861 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     1921 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1981 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2041 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2101 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2161 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2221 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2281 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2341 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2401 tgctgcttgc aaacnaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2461 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2521 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2581 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2701 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2761 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2821 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2881 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2941 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3001 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3061 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3121 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3181 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3241 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     3301 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3361 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3421 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     3481 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     3541 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     3601 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     3661 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     3721 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     3781 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     3841 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     3901 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     3961 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     4021 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt
     4081 gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt
     4141 cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag
     4201 gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg
     4261 gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc
     4321 gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta
     4381 agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc
     4441 atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag
     4501 gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggcagccatg
     4561 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     4621 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     4681 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     4741 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     4801 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc cttatgcgac
     4861 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     4921 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     4981 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5041 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5101 cgtccggcgt agagaatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg
     5161 gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg
     5221 agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga
     5281 ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc
     5341 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
     5401 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
     5461 aacttatcga tgataagctg tcaaacatga gaattgagct c
//
