LOCUS       sequence-414748-        7999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal TinyERRE-tagged transferrin receptor 1 (tag on 
            extracellular domain)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7999)
  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 7999)
  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-TinyERRE
FEATURES             Location/Qualifiers
     source          1..7999
                     /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(3552..3570)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3553..3570)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3590..3607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3596..3820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3866..4294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3953..3972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4163..4184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4303..4323)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4420..4629
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4441..4637
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4488..4623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4550..4569
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4704..5498
                     /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(4758..4777)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5368..5387
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5672..5793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5709..5728)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5763..5782
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5842..5858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5842..5858)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5855..5877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5866..5882
                     /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(5890..5896,5897..5914,5915..5920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5935..5956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6073..6090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6244..6832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6324..6343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7003..7794,7795..7863))
                     /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     7626..7645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7864..7968)
                     /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 ggtctcgaaa tcgaagttgc gttcctggaa cgtcgtgtta ccgcgctgcg tacccgtaac
     3361 gcggaactgc gtcaggaagt tcagcgtctg gaaaacgaat acggtccgct gaccggttaa
     3421 atgtaacatg cataattaaa taagagcagg gtagtctgtt tctaaaaaaa aaaaaaaaaa
     3481 aaaaaaaaaa ggaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
     3541 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3601 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3661 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3721 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3781 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3841 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3901 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3961 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4021 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4081 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4141 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4201 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4261 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4381 agtcagcaac catagtcccg cccctcmcwc cccccccasc aggcagaagt atgcaaagca
     4441 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4501 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4561 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4621 gcctaggctt ttgcaaaaag ctcctgggag cttgtatatc cattttcgga tctgatcaag
     4681 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4741 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4801 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4861 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4921 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4981 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5041 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5101 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5161 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5221 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5281 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5341 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5401 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5461 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5521 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5581 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5641 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5701 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5761 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5821 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5881 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5941 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6001 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7981 aaaagtgcca cctgacgtc
//
