LOCUS       sequence-377037-        7944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MiniE genetically encoded tag on Transferrin Receptor 1 (TfR1) to be
            used with MiniR labeled with a reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7944)
  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 7944)
  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-miniE
FEATURES             Location/Qualifiers
     source          1..7944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(158..177)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        349..728
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        729..932
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     883..903
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     977..996
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        977..995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3683..3701)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3684..3701)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3721..3738)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3727..3951
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3997..4425
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4084..4103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4294..4315
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4434..4454)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4478..4688
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4500..4696
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4547..4682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4763..5557
                     /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(4817..4836)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5427..5446
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5731..5852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5768..5787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5822..5841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5901..5917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5901..5917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5914..5936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5925..5941
                     /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(5949..5955,5956..5973,5974..5979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5994..6015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6132..6149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6303..6891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6383..6402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7062..7853,7854..7922))
                     /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     7685..7704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7923..7944,1..83))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
       61 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
      121 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
      181 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
      241 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
      301 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      361 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      421 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      481 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      721 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      781 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      841 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      901 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      961 actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg taccgagctc
     1021 ggatccacta gtaacggccg ccagtgtgct ggaattccgc ggactgtaga ggcgcttcct
     1081 agtactccct tgtagcagct gagaatgatg gatcaagcca gatcagcatt ctctaacttg
     1141 tttggtgggg aaccattgtc atacacccgg tttagccttg ctcggcaagt agatggagat
     1201 aacagtcatg tggagatgaa actggctgca gatgaagaag aaaatgccga caataacatg
     1261 aaggctagtg tcagaaaacc caagaggttt aatggaagac tctgctttgc agctattgca
     1321 ctagtcattt tcttcttgat tggattcatg agtggctacc tgggctattg taagcgtgta
     1381 gaacaaaaag aggagtgtgt gaaactggct gaaacggagg agacagacaa gtcagaaacc
     1441 atggaaacag aggatgttcc tacatcatct cgcttatatt gggcagacct caaaacactg
     1501 ttgtcagaga agttgaactc catagagttt gctgacacca tcaagcagct gagccagaat
     1561 acatacactc ctcgtgaggc tggatctcaa aaagatgaaa gtcttgccta ttatattgaa
     1621 aatcagttcc atgaatttaa attcagcaaa gtctggcgag atgaacacta tgtgaagatt
     1681 caagtgaaaa gcagcattgg tcaaaacatg gtgaccatag tgcagtcaaa tggtaactta
     1741 gacccagtgg agtctcccga gggttatgtg gcattcagta aacctacaga agtttctggt
     1801 aaactggtcc atgctaattt tggcactaaa aaggactttg aagaactaag ttattctgtg
     1861 aatggatctt tagtgattgt tagagcaggg gaaattactt ttgcagaaaa ggttgcaaat
     1921 gcccaaagct ttaatgcaat tggtgtcctc atatacatgg acaagaataa attccccgtt
     1981 gttgaggcag accttgcact ctttggacat gctcatctag gaactggtga tccatacaca
     2041 cctggctttc cttctttcaa tcatactcag tttccgccat ctcagtcatc agggttgcct
     2101 aatatacctg tgcaaacaat ctcaagagct gctgcagaaa agctatttgg aaaaatggaa
     2161 ggaagctgtc ctgctagatg gaacatagat tcttcatgta agctggaact ttcacagaat
     2221 caaaatgtga agctcattgt gaaaaacgta ctgaaagaaa gaagaatact taacatcttt
     2281 ggagttatta aaggttatga ggaaccagac cgttatgttg tagtaggagc ccagagagac
     2341 gctttgggtg ctggtgttgc ggcgaagtcc agtgtgggaa caggtcttct gttgaaactt
     2401 gcccaagtat tctcagatat gatttcaaaa gatggattta gacccagcag aagtataatc
     2461 tttgccagct ggactgcagg cgactttgga gctgttggtg ccactgagtg gttggaggga
     2521 tacctttcat ctttgcattt aaaagctttc acttatatta atttggataa agttgtcctt
     2581 ggtactagta acttcaaagt ttctgccagc cccttattat atacacttat gggaaagata
     2641 atgcaagatg taaagcatcc agttgatgga aaatctctat atagagacag caattggatt
     2701 agcaaagttg agaaactttc ctttgacaat gctgcatatc ctttccttgc atattctgga
     2761 atcccagcag tttctttttg tttttgtgag gatgcagact atccttattt gggcactaga
     2821 ttggatacct atgaggcatt gactcagaaa gttcctcagc tcaaccaaat ggttcgtaca
     2881 gcagcggaag tggctggtca gctcattatt aaacttaccc atgacgttga attgaacctg
     2941 gactatgaga tgtataacag caaactactg tcatttatga aggatctgaa ccagttcaaa
     3001 acagatatca gggatatggg tctaagtcta cagtggctgt attccgctcg tggagactac
     3061 ttccgtgcta cttctagact aacaactgat tttcataatg ctgagaaaac aaacagattt
     3121 gtcatgaggg aaatcaatga tcgtattatg aaagtggagt atcacttcct gtcgccctat
     3181 gtatctccaa gagagtctcc tttccgacat atcttctggg gctctggctc tcacactctc
     3241 tcagctttag tggagaactt gaagcttcgt caaaaaaata ttactgcttt taatgaaacc
     3301 ctcttcagaa accagttggc cctggctact tggactattc agggagtcgc aaatgccctc
     3361 tctggtgaca tttggaatat tgacaatgag tttggtagcg gcagcggtag caccggtatg
     3421 ctggagatcg aggccgcctt cctcgagagg gagaacaccg ccctggagac cagggtggcc
     3481 gagctgaggc agagggtgca gaggctgagg aacgagtacg gccctctggg aggcggaacc
     3541 ggttaaatgt aacatgcata attaaataag agcagggtag tctgtttcta gaaaaaaaaa
     3601 aaaaaaaaaa aaaaaaaaaa aggaattctg cagatatcca tcacactggc ggccgctcga
     3661 gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag
     3721 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3781 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3841 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3901 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3961 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4021 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4081 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4141 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4201 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4261 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4321 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4381 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4441 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4501 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4561 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4621 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4681 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4741 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4801 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4861 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4921 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4981 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5041 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5101 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5161 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5221 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5281 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5341 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5401 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5461 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5521 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5581 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5641 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5701 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5821 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5881 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6721 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7921 atactcttcc tttttcaata ttat
//
