LOCUS       sequence-393648-        4168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for N-terminal HiBIT-tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4168)
  AUTHORS   Schwalm MP, Kramer A, Dolle A, Weckesser J, Yu X, Jin J, Saxena K, 
            Knapp S
  TITLE     Tracking the PROTAC degradation pathway in living cells highlights 
            the importance of ternary complex measurement for PROTAC 
            optimization.
  JOURNAL   Cell Chem Biol. 2023 Jun 20:S2451-9456(23)00157-5. doi: 
            10.1016/j.chembiol.2023.06.002.
  PUBMED    37354907
REFERENCE   2  (bases 1 to 4168)
  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: "Cell Chem 
            Biol. 2023 Jun 20:S2451-9456(23)00157-5. doi: 
            10.1016/j.chembiol.2023.06.002."
COMMENT     Sequence Label: pF-N-HiBiT-template
FEATURES             Location/Qualifiers
     source          1..4168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..573
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     474..493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     690..707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        1152..1531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1532..1735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1686..1706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1871..2003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1986..2005
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     2047..2066
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2047..2065
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2261..2279)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2316..2437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2327..2346)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2381..2400
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2682..3039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2682..2873
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2705..2725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2890..3025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3054..3101
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3133..3927
                     /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(3187..3206)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3797..3816
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3991..4039
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg
       61 ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa
      121 ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt
      181 ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg
      241 ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg
      361 cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg
      421 acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac
      481 gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg
      541 tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt
      601 ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt
      661 ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata
      721 tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag
      781 cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc
      841 tgatgtccgg cggtgctttt gccgttacgc accaccccgt catgagatta cgcgttttag
      901 cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg
      961 agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag atcttcaata
     1021 ttggccatta gccatattat tcattggtta tatagcataa atcaatattg gctattggcc
     1081 attgcatacg ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg
     1141 accgccatgt tggcattgat tattgactag ttattaatag taatcaatta cggggtcatt
     1201 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     1261 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     1321 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     1381 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
     1441 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
     1501 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
     1561 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     1621 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
     1681 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     1741 agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt aaattgctaa
     1801 cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt ggtcgtgagg
     1861 cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     1921 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
     1981 ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct cttaaggcta
     2041 gagtattaat acgactcact atagggctag cgctcaccat ggtcgtgagc ggctggcggc
     2101 tgttcaagaa gattagcaac aggatcaggg gcagcagcgg cggcagcagc taaacgaatt
     2161 cgggctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc tgatccggct
     2221 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     2281 taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2341 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2401 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2461 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     2521 atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     2581 tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg
     2641 cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag
     2701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2941 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaatta actgttgaca
     3061 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     3121 aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3181 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3241 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3301 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3361 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3421 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3481 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3541 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3601 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3661 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3721 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3781 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3841 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3901 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     3961 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     4021 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc gctcatgcga tgataagctg
     4081 tcaaacatga gaattacaac ttatatcgta tggggctgac ttcaggtgct acatttgaag
     4141 agataaattg cactgaaatc tagaaata
//
