LOCUS       sequence-357271-        5397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to label proteins with the MiniE genetically encoded tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5397)
  AUTHORS   Doh JK, Tobin SJ, Beatty KE
  TITLE     MiniVIPER Is a Peptide Tag for Imaging and Translocating Proteins in
            Cells.
  JOURNAL   Biochemistry. 2020 Aug 25;59(33):3051-3059. doi: 
            10.1021/acs.biochem.0c00526. Epub 2020 Aug 17.
  PUBMED    32786411
REFERENCE   2  (bases 1 to 5397)
  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/acs.biochem.0c00526"; journalName: "Biochemistry"; date: 
            "2020-08-25- 25"; volume: "59"; issue: "33"; pages: "3051-3059"
COMMENT     Sequence Label: pET28b-miniR-DBCO
FEATURES             Location/Qualifiers
     source          1..5397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1313..1504
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1498..1520)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1606..1745
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1661..1680
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1760..1777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1931..2519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2011..2030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2630..2649)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2641..3456
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2641..3456
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2714..2733)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3548..4003)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3685..3706)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3897..3916
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4040..4087)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4083..4101
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4154..4171)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4352..4374
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4389..4413
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(4413..4432)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4414..4432)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4516..4535)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4741..4818
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4819..5397,1..504)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4838..4857)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
       61 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
      121 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
      181 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
      241 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
      301 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
      361 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
      421 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      481 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
      541 ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
      601 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
      661 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
      721 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
      781 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
      841 ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
      901 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
      961 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     1021 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     1081 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     1141 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     1201 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     1261 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     1321 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     1381 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     1441 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     1501 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     1561 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     1621 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     1681 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     1741 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1801 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1861 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1921 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1981 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2041 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2101 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2161 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2221 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2281 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2341 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2401 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2461 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2521 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2581 ggattttggt catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2641 atgagccata ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat
     2701 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     2761 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     2821 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2881 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2941 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     3001 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     3061 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     3121 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     3181 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     3241 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     3301 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     3361 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     3421 ttgcagtttc atttgatgct cgatgagttt ttctaagaat taattcatga gcggatacat
     3481 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3541 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3601 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3661 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3721 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3781 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3841 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3901 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3961 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccaatccgg
     4021 atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     4081 atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag
     4141 cagccggatc tcagtggtgg tggtggtggt gctcgagacc gtaaacccac gggtagcaca
     4201 gaccccatgc cgcagcacca ccacccagcg gaccgtaacg gttttccaga cgctgaactt
     4261 cctgttccag ttccgcaact tcggtacgca gcgcggtgtt acgctgacgc aggaacgcaa
     4321 cacggatttc gagagaagaa cccatggtat atctccttct taaagttaaa caaaattatt
     4381 tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg
     4441 ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     4501 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     4561 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     4621 tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca
     4681 acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgagatcccg
     4741 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     4801 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     4861 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     4921 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
     4981 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
     5041 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
     5101 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
     5161 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
     5221 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
     5281 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
     5341 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcg
//
