LOCUS       sequence-357263-        7946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MiniR genetically encoded tag on Transferrin Receptor 1 (TfR1) to be
            used with MiniE labeled with a reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7946)
  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 7946)
  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: pcDNA TfR1-miniR
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(151..170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        342..721
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        722..925
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     876..896
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     970..989
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        970..988
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3678..3696)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3679..3696)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3716..3733)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3722..3946
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3992..4420
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4079..4098)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4289..4310
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4429..4449)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4473..4683
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4495..4691
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4542..4677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4604..4623
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4758..5552
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4812..4831)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5422..5441
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5726..5847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5763..5782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5817..5836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5896..5912)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5896..5912)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5909..5931)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5920..5936
                     /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)."
     promoter        complement(join(5944..5950,5951..5968,5969..5974))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5989..6010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6127..6144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6298..6886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6378..6397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7057..7848,7849..7917))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7680..7699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7918..7946,1..76))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
       61 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
      121 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      181 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
      241 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
      301 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      361 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      421 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      481 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      541 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      601 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      661 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      721 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      781 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      841 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      901 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
      961 tatcgaaatt aatacgactc actataggga gacccaagct tggtaccgag ctcggatcca
     1021 ctagtaacgg ccgccagtgt gctggaattc cgcggactgt agaggcgctt cctagtactc
     1081 ccttgtagca gctgagaatg atggatcaag ccagatcagc attctctaac ttgtttggtg
     1141 gggaaccatt gtcatacacc cggtttagcc ttgctcggca agtagatgga gataacagtc
     1201 atgtggagat gaaactggct gcagatgaag aagaaaatgc cgacaataac atgaaggcta
     1261 gtgtcagaaa acccaagagg tttaatggaa gactctgctt tgcagctatt gcactagtca
     1321 ttttcttctt gattggattc atgagtggct acctgggcta ttgtaagcgt gtagaacaaa
     1381 aagaggagtg tgtgaaactg gctgaaacgg aggagacaga caagtcagaa accatggaaa
     1441 cagaggatgt tcctacatca tctcgcttat attgggcaga cctcaaaaca ctgttgtcag
     1501 agaagttgaa ctccatagag tttgctgaca ccatcaagca gctgagccag aatacataca
     1561 ctcctcgtga ggctggatct caaaaagatg aaagtcttgc ctattatatt gaaaatcagt
     1621 tccatgaatt taaattcagc aaagtctggc gagatgaaca ctatgtgaag attcaagtga
     1681 aaagcagcat tggtcaaaac atggtgacca tagtgcagtc aaatggtaac ttagacccag
     1741 tggagtctcc cgagggttat gtggcattca gtaaacctac agaagtttct ggtaaactgg
     1801 tccatgctaa ttttggcact aaaaaggact ttgaagaact aagttattct gtgaatggat
     1861 ctttagtgat tgttagagca ggggaaatta cttttgcaga aaaggttgca aatgcccaaa
     1921 gctttaatgc aattggtgtc ctcatataca tggacaagaa taaattcccc gttgttgagg
     1981 cagaccttgc actctttgga catgctcatc taggaactgg tgatccatac acacctggct
     2041 ttccttcttt caatcatact cagtttccgc catctcagtc atcagggttg cctaatatac
     2101 ctgtgcaaac aatctcaaga gctgctgcag aaaagctatt tggaaaaatg gaaggaagct
     2161 gtcctgctag atggaacata gattcttcat gtaagctgga actttcacag aatcaaaatg
     2221 tgaagctcat tgtgaaaaac gtactgaaag aaagaagaat acttaacatc tttggagtta
     2281 ttaaaggtta tgaggaacca gaccgttatg ttgtagtagg agcccagaga gacgctttgg
     2341 gtgctggtgt tgcggcgaag tccagtgtgg gaacaggtct tctgttgaaa cttgcccaag
     2401 tattctcaga tatgatttca aaagatggat ttagacccag cagaagtata atctttgcca
     2461 gctggactgc aggcgacttt ggagctgttg gtgccactga gtggttggag ggataccttt
     2521 catctttgca tttaaaagct ttcacttata ttaatttgga taaagttgtc cttggtacta
     2581 gtaacttcaa agtttctgcc agccccttat tatatacact tatgggaaag ataatgcaag
     2641 atgtaaagca tccagttgat ggaaaatctc tatatagaga cagcaattgg attagcaaag
     2701 ttgagaaact ttcctttgac aatgctgcat atcctttcct tgcatattct ggaatcccag
     2761 cagtttcttt ttgtttttgt gaggatgcag actatcctta tttgggcact agattggata
     2821 cctatgaggc attgactcag aaagttcctc agctcaacca aatggttcgt acagcagcgg
     2881 aagtggctgg tcagctcatt attaaactta cccatgacgt tgaattgaac ctggactatg
     2941 agatgtataa cagcaaacta ctgtcattta tgaaggatct gaaccagttc aaaacagata
     3001 tcagggatat gggtctaagt ctacagtggc tgtattccgc tcgtggagac tacttccgtg
     3061 ctacttctag actaacaact gattttcata atgctgagaa aacaaacaga tttgtcatga
     3121 gggaaatcaa tgatcgtatt atgaaagtgg agtatcactt cctgtcgccc tatgtatctc
     3181 caagagagtc tcctttccga catatcttct ggggctctgg ctctcacact ctctcagctt
     3241 tagtggagaa cttgaagctt cgtcaaaaaa atattactgc ttttaatgaa accctcttca
     3301 gaaaccagtt ggccctggct acttggacta ttcagggagt cgcaaatgcc ctctctggtg
     3361 acatttggaa tattgacaat gagtttggta gcggcagcgg tagcaccggt atgggttctt
     3421 ctctcgaaat ccgtgttgcg ttcctgcgtc agcgtaacac cgcgctgcgt accgaagttg
     3481 cggaactgga acaggaagtt cagcgtctgg aaaaccgtta cggtccgctg accggttaaa
     3541 tgtaacatgc ataattaaat aagagcaggg tagtctgttt ctagaaaaaa aaaaaaaaaa
     3601 aaaaaaaaaa aaaaaaggaa ttctgcagat atccatcaca ctggcggccg ctcgagcatg
     3661 catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg
     3721 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3781 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3841 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3901 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3961 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4021 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4081 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4141 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4201 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4261 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4321 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4381 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4441 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4501 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4561 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4621 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4681 gcttttgcaa aaagctcctg ggagcttgta tatccatttt cggatctgat caagagacag
     4741 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4801 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4861 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4921 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4981 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5041 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5101 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5161 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5221 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5281 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5341 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5401 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5461 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5521 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5581 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5641 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5701 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5821 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5881 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5941 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6001 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6061 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6121 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6181 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6241 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6301 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6361 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6421 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6481 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6541 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6601 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6661 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6721 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6781 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6841 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6901 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6961 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7021 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7081 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7141 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7201 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7261 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7321 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7381 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7441 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7501 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7561 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7621 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7681 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7741 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7801 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7861 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7921 cttccttttt caatattatt gaagca
//
