LOCUS       sequence-213810-        5253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses BC2-tag-Ypet-FtnA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5253)
  AUTHORS   Virant D, Traenkle B, Maier J, Kaiser PD, Bodenhofer M, Schmees C, 
            Vojnovic I, Pisak-Lukats B, Endesfelder U, Rothbauer U
  TITLE     A peptide tag-specific nanobody enables high-quality labeling for 
            dSTORM imaging.
  JOURNAL   Nat Commun. 2018 Mar 2;9(1):930. doi: 10.1038/s41467-018-03191-2.
  PUBMED    29500346
REFERENCE   2  (bases 1 to 5253)
  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"; journalName: "Nat 
            Commun."; date: "2018-03-2"; volume: "9(1):930. doi"; pages: " 
            10.1038/s41467-018-03191-2"
COMMENT     Sequence Label: pBC2T-Ypet-FtnA
FEATURES             Location/Qualifiers
     source          1..5253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(704..706,707..1417)
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLFKDPNEKRDHMVLLE
                     FLTAAGITEGMNELYK"
     primer_bind     complement(1007..1026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2099..2220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2136..2155)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2190..2209
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2227..2682)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2364..2385)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2576..2595
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2709..2813
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2815..3172
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2815..3006
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2838..2858)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3023..3158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3207..4001
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3207..4001
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3261..3280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3871..3890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4189..4208)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4233..4280
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4609..5197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5098..5117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc caccggtcgc
      661 caccatgcct gatcgcaagc gcgccgtgag ccattggcag caggtgagca aaggcgaaga
      721 gctgttcacc ggcgtggtgc ccatcctggt ggagctggac ggcgacgtga acggccacaa
      781 gttcagcgtg agcggcgagg gcgagggcga cgccacctac ggcaagctga ccctgaagct
      841 gctgtgcacc accggcaagc tgcccgtgcc ctggcccacc ctggtgacca ccctgggcta
      901 cggcgtgcag tgcttcgccc ggtaccccga ccacatgaag cagcacgact tcttcaagag
      961 cgccatgccc gagggctacg tgcaggagcg gaccatcttc ttcaaggacg acggcaacta
     1021 caagacccgg gccgaggtga agttcgaggg cgacaccctg gtgaaccgga tcgagctgaa
     1081 gggcatcgac ttcaaggagg acggcaacat cctgggccac aagctggagt acaactacaa
     1141 cagccacaac gtgtacatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     1201 gatccggcac aacatcgagg acggcggcgt gcagctggcc gaccactacc agcagaacac
     1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagagcaa
     1321 gctgttcaag gaccccaacg agaagcggga ccacatggtg ctgctggagt tcctgaccgc
     1381 cgccggcatc accgagggca tgaacgagct ctataaggct agcggagcat ccgcaggaat
     1441 gctgaaacca gaaatgattg aaaaacttaa tgagcagatg aacctggaac tgtactcttc
     1501 actgctttat cagcaaatga gcgcctggtg cagctatcat accttcgaag gtgctgccgc
     1561 gttcctgcgc cgtcacgccc aggaagagat gacgcatatg cagcgtctgt ttgattacct
     1621 gactgatacc ggcaatttac cgcgtattaa taccgttgaa tctccgtttg ctgaatattc
     1681 ctcacttgat gaattattcc aggaaaccta taaacacgaa caattaatca cccagaaaat
     1741 taacgaactg gctcatgctg caatgaccaa tcaggactac ccaacattta atttcctgca
     1801 atggtatgtt tctgagcagc atgaagaaga gaaactgttc aaatcgatta ttgataaatt
     1861 aagcctggca ggcaaaagcg gcgaaggtct gtattttatc gacaaagaac tctctaccct
     1921 cgacacacaa aactaacggc cgcatcgata acaccgcaaa tgcttctagg gatccaccgg
     1981 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2041 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2101 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2161 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2221 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2281 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2341 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2401 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2461 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2521 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2581 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2641 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2701 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2761 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2821 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2881 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2941 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3001 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3061 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3121 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3181 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3241 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3301 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3361 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3421 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3481 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3541 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3601 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3661 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3721 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3781 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3841 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3901 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3961 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4021 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4081 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4141 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4201 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4261 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4321 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4381 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4441 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4501 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4561 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4621 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4681 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4741 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4801 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4861 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4921 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4981 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5041 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5101 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5161 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5221 acggttcctg gccttttgct ggccttttgc tca
//
