LOCUS       sequence-414751-        7973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal PunyEEER-tagged transferrin receptor 1 (tag on 
            extracellular domain)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7973)
  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 7973)
  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-PunyEEER
FEATURES             Location/Qualifiers
     source          1..7973
                     /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(3526..3544)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3527..3544)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3564..3581)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3570..3794
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3840..4268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3927..3946)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4137..4158
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4277..4297)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4394..4603
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4415..4611
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4462..4597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4524..4543
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4678..5472
                     /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(4732..4751)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5342..5361
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5646..5767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5683..5702)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5737..5756
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5816..5832)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5816..5832)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5829..5851)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5840..5856
                     /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(5864..5870,5871..5888,5889..5894))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5909..5930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6047..6064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6218..6806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6298..6317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6977..7768,7769..7837))
                     /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     7600..7619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7838..7942)
                     /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 gtcatgcgga 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 ggtctggaga ttgaaaacgc ctttctggag cgtgaggtga ccgccctgga gaccgaggtg
     3361 cagcgcctgg aacagcgtaa ccagcgtctg cgcaaccgtt atggtccact gaccggttaa
     3421 atgtaacatg cataattaaa taagagcagg gtagtctgtt tctaggaatt ctgcagatat
     3481 ccatcacact ggcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct
     3541 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3601 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3661 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3721 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3781 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3841 cgcgccctgt agctgcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3901 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3961 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4021 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4081 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4141 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4201 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4261 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4381 acwccccccc cascaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4441 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4561 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctg
     4621 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4681 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4741 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4801 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4861 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4921 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4981 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5041 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5101 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5161 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5221 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5281 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5341 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5401 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5461 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5521 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5581 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5641 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5701 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5761 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5821 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5881 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5941 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6001 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6061 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6121 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6181 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6241 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6301 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6361 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6421 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6481 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6541 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6601 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6661 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6721 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6781 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6841 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6901 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6961 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7021 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7081 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7141 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7201 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7261 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7321 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7381 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7441 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7501 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7561 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7621 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7681 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7741 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7801 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7861 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7921 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
