LOCUS       sequence-414756-        5244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TREER genetically-encoded tag introduced before actin; to be used 
            with TinyERRE probe peptide..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5244)
  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 5244)
  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: TREER-Actin
FEATURES             Location/Qualifiers
     source          1..5244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..291
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(293..397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      424..879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(511..530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     721..742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    886..1007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(897..916)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     951..970
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2471..2674)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2500..2520)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2675..2978
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3153..3741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3233..3252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4070..4117
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4142..4161
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4349..5143)
                     /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             complement(4349..5143)
                     /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(4460..4479)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5070..5089
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5178..5244,1..291))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5192..5244,1..83)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc
      301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      421 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      841 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cgcgttaaga tacattgatg
      901 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      961 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1021 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1081 acctctacaa atgtggtatg gctgattatg atcagttatc tagatccggt ggatccctag
     1141 aagcatttgc ggtggacgat ggaggggccg gactcgtcat actcctgctt gctgatccac
     1201 atctgctgga aggtggacag cgaggccagg atggagccgc cgatccacac ggagtacttg
     1261 cgctcaggag gagcaatgat cttgatcttc attgtgctgg gtgccagggc agtgatctcc
     1321 ttctgcatcc tgtcggcaat gccagggtac atggtggtgc cgccagacag cactgtgttg
     1381 gcgtacaggt ctttgcggat gtccacgtca cacttcatga tggagttgaa ggtagtttcg
     1441 tggatgccac aggactccat gcccaggaag gaaggctgga agagtgcctc agggcagcgg
     1501 aaccgctcat tgccaatggt gatgacctgg ccgtcaggca gctcgtagct cttctccagg
     1561 gaggagctgg aagcagccgt ggccatctct tgctcgaagt ccagggcgac gtagcacagc
     1621 ttctccttaa tgtcacgcac gatttcccgc tcggccgtgg tggtgaagct gtagccgcgc
     1681 tcggtgagga tcttcatgag gtagtcagtc aggtcccggc cagccaggtc cagacgcagg
     1741 atggcatggg ggagggcata cccctcgtag atgggcacag tgtgggtgac cccgtcaccg
     1801 gagtccatca cgatgccagt ggtacggcca gaggcgtaca gggatagcac agcctggata
     1861 gcaacgtaca tggctggggt gttgaaggtc tcaaacatga tctgggtcat cttctcgcgg
     1921 ttggccttgg ggttcagggg ggcctcggtc agcagcacgg ggtgctcctc gggagccaca
     1981 cgcagctcat tgtagaaggt gtggtgccag attttctcca tgtcgtccca gttggtgacg
     2041 atgccgtgct cgatggggta cttcagggtg aggatgcctc tcttgctctg ggcctcgtcg
     2101 cccacatagg aatccttctg acccatgccc accatcacgc cctggtgcct ggggcgcccc
     2161 acgatggagg ggaagacggc ccggggggca tcgtcgcccg cgaagccggc cttgcacatg
     2221 ccggagccgt tgtcgacgac gagcgcggcg atatcatcat cgcttccgct tccgccggat
     2281 gcgctgcctc caccgctgcc agatctgcct ccacccagcg gaccgtaacg gttacgcaga
     2341 cgctgaacac gctgttccag ttccgcgttt tcggtttcca gcgcggtgac ttcctgacgc
     2401 aggaacgcaa cacggatttc cagcatggtg gcgaccggta gcgctagcgg atctgacggt
     2461 tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     2521 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     2581 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     2641 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     2701 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     2761 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     2821 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     2881 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     2941 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     3001 gaactaatga ccccgtaatt gattactatt aataactaat gcatggcggt aatacggtta
     3061 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3121 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3181 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3241 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3301 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3361 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3421 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3481 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3541 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3601 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3661 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3721 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3781 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3841 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct
     3901 gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
     3961 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
     4021 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
     4081 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
     4141 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     4201 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     4261 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     4321 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     4381 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     4441 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     4501 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     4561 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     4621 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4681 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     4741 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4801 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4861 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     4921 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     4981 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     5041 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     5101 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     5161 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     5221 tggaatagct cagaggccga ggcg
//
