LOCUS       sequence-414752-        7975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal PuRRRE-tagged transferrin receptor 1 (tag on 
            extracellular domain)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7975)
  AUTHORS   Suyama A, Devlin KL, Macias-Contreras M, Doh JK, Shinde U, Beatty KE
  TITLE     Orthogonal Versatile Interacting Peptide Tags for Imaging Cellular 
            Proteins.
  JOURNAL   Biochemistry. 2023 Jun 6;62(11):1735-1743. doi: 
            10.1021/acs.biochem.2c00712. Epub 2023 May 11.
  PUBMED    37167569
REFERENCE   2  (bases 1 to 7975)
  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.2c00712"; journalName: "Biochemistry"; date: 
            "2023-06-6- 6"; volume: "62"; issue: "11"; pages: "1735-1743"
COMMENT     Sequence Label: pcDNA TfR1-PuRRRE
FEATURES             Location/Qualifiers
     source          1..7975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3528..3546)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3529..3546)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3566..3583)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3572..3796
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3842..4270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3929..3948)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4139..4160
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4279..4299)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4395..4605
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4417..4613
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4464..4599
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4526..4545
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4680..5474
                     /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(4734..4753)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5344..5363
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5648..5769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5685..5704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5739..5758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5818..5834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5818..5834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5831..5853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5842..5858
                     /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(5866..5872,5873..5890,5891..5896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5911..5932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6049..6066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6220..6808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6300..6319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6979..7770,7771..7839))
                     /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     7602..7621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7840..7944)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttccgcggac tgtagaggcg
      961 cttcctagta ctcccttgta gcagctgaga atgatggatc aagccagatc agcattctct
     1021 aacttgtttg gtggggaacc attgtcatac acccggttta gccttgctcg gcaagtagat
     1081 ggagataaca gtcatgtgga gatgaaactg gctgcagatg aagaagaaaa tgccgacaat
     1141 aacatgaagg ctagtgtcag aaaacccaag aggtttaatg gaagactctg ctttgcagct
     1201 attgcactag tcattttctt cttgattgga ttcatgagtg gctacctggg ctattgtaag
     1261 cgtgtagaac aaaaagagga gtgtgtgaaa ctggctgaaa cggaggagac agacaagtca
     1321 gaaaccatgg aaacagagga tgttcctaca tcatctcgct tatattgggc agacctcaaa
     1381 acactgttgt cagagaagtt gaactccata gagtttgctg acaccatcaa gcagctgagc
     1441 cagaatacat acactcctcg tgaggctgga tctcaaaaag atgaaagtct tgcctattat
     1501 attgaaaatc agttccatga atttaaattc agcaaagtct ggcgagatga acactatgtg
     1561 aagattcaag tgaaaagcag cattggtcaa aacatggtga ccatagtgca gtcaaatggt
     1621 aacttagacc cagtggagtc tcccgagggt tatgtggcat tcagtaaacc tacagaagtt
     1681 tctggtaaac tggtccatgc taattttggc actaaaaagg actttgaaga actaagttat
     1741 tctgtgaatg gatctttagt gattgttaga gcaggggaaa ttacttttgc agaaaaggtt
     1801 gcaaatgccc aaagctttaa tgcaattggt gtcctcatat acatggacaa gaataaattc
     1861 cccgttgttg aggcagacct tgcactcttt ggacatgctc atctaggaac tggtgatcca
     1921 tacacacctg gctttccttc tttcaatcat actcagtttc cgccatctca gtcatcaggg
     1981 ttgcctaata tacctgtgca aacaatctca agagctgctg cagaaaagct atttggaaaa
     2041 atggaaggaa gctgtcctgc tagatggaac atagattctt catgtaagct ggaactttca
     2101 cagaatcaaa atgtgaagct cattgtgaaa aacgtactga aagaaagaag aatacttaac
     2161 atctttggag ttattaaagg ttatgaggaa ccagaccgtt atgttgtagt aggagcccag
     2221 agagacgctt tgggtgctgg tgttgcggcg aagtccagtg tgggaacagg tcttctgttg
     2281 aaacttgccc aagtattctc agatatgatt tcaaaagatg gatttagacc cagcagaagt
     2341 ataatctttg ccagctggac tgcaggcgac tttggagctg ttggtgccac tgagtggttg
     2401 gagggatacc tttcatcttt gcatttaaaa gctttcactt atattaattt ggataaagtt
     2461 gtccttggta ctagtaactt caaagtttct gccagcccct tattatatac acttatggga
     2521 aagataatgc aagatgtaaa gcatccagtt gatggaaaat ctctatatag agacagcaat
     2581 tggattagca aagttgagaa actttccttt gacaatgctg catatccttt ccttgcatat
     2641 tctggaatcc cagcagtttc tttttgtttt tgtgaggatg cagactatcc ttatttgggc
     2701 actagattgg atacctatga ggcattgact cagaaagttc ctcagctcaa ccaaatggtt
     2761 cgtacagcag cggaagtggc tggtcagctc attattaaac ttacccatga cgttgaattg
     2821 aacctggact atgagatgta taacagcaaa ctactgtcat ttatgaagga tctgaaccag
     2881 ttcaaaacag atatcaggga tatgggtcta agtctacagt ggctgtattc cgctcgtgga
     2941 gactacttcc gtgctacttc tagactaaca actgattttc ataatgctga gaaaacaaac
     3001 agatttgtca tgagggaaat caatgatcgt attatgaaag tggagtatca cttcctgtcg
     3061 ccctatgtat ctccaagaga gtctcctttc cgacatatct tctggggctc tggctctcac
     3121 actctctcag ctttagtgga gaacttgaag cttcgtcaaa aaaatattac tgcttttaat
     3181 gaaaccctct tcagaaacca gttggccctg gctacttgga ctattcaggg agtcgcaaat
     3241 gccctctctg gtgacatttg gaatattgac aatgagtttg gtagcggcag cggtagcacc
     3301 ggtctggaaa tccgtaacgc cttcctccgt cagcgtgtta ccgcgctgcg tacccgcgtt
     3361 tctgaactgc gtcaggaaaa ctccgaactg gaaaacgaat acggtccgct gtaaaccggt
     3421 taaatgtaac atgcataatt aaataagagc agggtagtct gtttctagaa ttctgcagat
     3481 atccatcaca ctggcggccg ctcgagcatg catctagagg gccctattct atagtgtcac
     3541 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3601 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3661 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3721 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3781 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3841 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3901 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3961 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4021 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4081 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4141 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4201 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4261 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4381 tcccwccccc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4501 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4561 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4621 tgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4681 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4741 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4801 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4861 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4921 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4981 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5041 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5101 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5161 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5221 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5281 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5341 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5401 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5461 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5521 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5581 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5641 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5701 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5761 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5821 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5881 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5941 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6001 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6061 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6121 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6661 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6781 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6901 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6961 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7021 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7081 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7141 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7201 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7261 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7321 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7381 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7441 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7501 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7561 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7621 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7681 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7741 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7801 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7861 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7921 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
