LOCUS       sequence-357327-        8862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AtAFB2 (auxin-inducible degron system) fused to mTagBFP2 
            (lentiviral plasmid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8862)
  AUTHORS   Gemperle J, Harrison TS, Flett C, Adamson AD, Caswell PT
  TITLE     On demand expression control of endogenous genes with DExCon, 
            DExogron and LUXon reveals differential dynamics of Rab11 family 
            members.
  JOURNAL   Elife. 2022 Jun 16;11. pii: 76651. doi: 10.7554/eLife.76651.
  PUBMED    35708998
REFERENCE   2  (bases 1 to 8862)
  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: "Elife."; 
            date: "2022-06-16"; volume: "11. pii: 76651. doi"; pages: " 
            10.7554/eLife.76651"
COMMENT     Sequence Label: pCDH-AtAFB2-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..8862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             41..274
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    346..467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(383..402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     437..456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      507..642
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(675..691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(675..691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(688..710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    699..715
                     /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(723..729,730..747,748..753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    768..789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(906..923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1077..1665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(1836..2627,2628..2696))
                     /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     2459..2478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2697..2801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2869..2887
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2925..2947)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3047..3066
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3260..3282
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3274..3291
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3275..3291
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3306..3532
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3533..3713
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3760..3885
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4378..4611
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4796..4840
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5107..5223
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5266..5477
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5490..5758
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             7525..8235
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    8241..8829
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8294..8314)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8712..8723)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
       61 caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
      121 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      181 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      241 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
      301 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
      361 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
      421 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
      481 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
      541 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
      601 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacc
      661 aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
      721 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
      781 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
      841 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
      901 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1801 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1861 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1921 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1981 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2041 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2101 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2161 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2221 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2281 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2341 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2401 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     2461 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2521 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2581 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2641 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2701 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2761 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2821 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2881 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2941 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     3001 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     3061 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     3121 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     3181 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     3241 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
     3301 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
     3361 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
     3421 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
     3481 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
     3541 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     3601 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     3661 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
     3721 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
     3781 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
     3841 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
     3901 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
     3961 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
     4021 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
     4081 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
     4141 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
     4201 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
     4261 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     4321 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     4381 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     4441 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     4501 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     4561 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     4621 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     4681 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     4741 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     4801 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     4861 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     4921 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     4981 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     5041 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     5101 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     5161 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat
     5221 tttatcgata ctagtggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     5281 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     5341 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     5401 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     5461 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     5521 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     5581 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     5641 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     5701 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     5761 agctgtgacc ggcgcctact ctagatgaac tactttcccg atgaggtcat tgagcacgtc
     5821 tttgatttcg tcacaagcca caaggatagg aacgccatta gcctggtctg taagagctgg
     5881 tacaagatcg agcggtattc cagacagaag gtgttcatcg gcaactgcta cgccatcaat
     5941 ccagagaggc tgctgcggag atttccctgt ctgaagagcc tgaccctgaa gggcaagccc
     6001 cacttcgccg actttaacct ggtgcctcac gagtggggag gattcgtgct gccatggatc
     6061 gaggccctgg cacggtccag agtgggcctg gaggagctga ggctgaagcg catggtggtg
     6121 acagacgagt ctctggagct gctgtctcgg agcttcgtga acttcaagag cctggtgctg
     6181 gtgtcctgcg agggctttac cacagatggc ctggcaagca tcgcagcaaa ctgtaggcac
     6241 ctgcgggacc tggacctgca ggagaatgag atcgacgatc acagaggcca gtggctgtcc
     6301 tgcttccccg atacctgtac cacactggtg acactgaact ttgcctgcct ggagggcgag
     6361 acaaatctgg tggccctgga gaggctggtg gcacgctctc ccaacctgaa gagcctgaag
     6421 ctgaataggg cagtgcctct ggacgcactg gcaagactga tggcatgcgc accacagatc
     6481 gtggatctgg gcgtgggctc ctacgagaac gaccctgatt ccgagtctta tctgaagctg
     6541 atggccgtga tcaagaagtg taccagcctg cgcagcctgt ccggcttcct ggaggcagca
     6601 cctcactgcc tgtccgcctt tcacccaatc tgtcacaacc tgacatccct gaatctgtct
     6661 tacgccgccg agatccacgg ctcccacctg atcaagctga tccagcactg caagaagctg
     6721 cagaggctgt ggatcctgga ctccatcggc gataagggcc tggaggtggt ggcctctacc
     6781 tgtaaggagc tgcaggagct gcgcgtgttc ccatctgacc tgctgggagg aggaaacacc
     6841 gcagtgacag aggagggcct ggtggcaatc tctgccggat gccccaagct gcacagcatc
     6901 ctgtattttt gtcagcagat gaccaacgcc gccctggtga cagtggccaa gaactgcccc
     6961 aatttcatca ggtttcgcct gtgcatcctg gagcccaata agcctgacca cgtgacatct
     7021 cagcctctgg atgagggctt cggcgccatc gtgaaggcct gcaagagcct gaggcgcctg
     7081 tctctgagcg gcctgctgac cgaccaggtg ttcctgtaca tcggcatgta tgccaaccag
     7141 ctggagatgc tgtctatcgc ctttgccggc gacacagata agggcatgct gtacgtgctg
     7201 aatggctgta agaagatgaa gaagctggag atcagagaca gcccttttgg cgataccgcc
     7261 ctgctggcag acgtgagcaa gtatgagaca atgcggagcc tgtggatgag ctcctgcgag
     7321 gtgaccctga gcggctgtaa gagactggcc gagaaggccc catggctgaa cgtggagatc
     7381 atcaacgaga atgacaacaa taggatggag gagaatggcc acgagggccg ccagaaggtg
     7441 gataagctgt atctgtatag gaccgtcgtc gggactcgga tggatgcccc cccctttgtc
     7501 tggattctgg gcggatctgg cgggatggtg tctaagggcg aagagctgat taaggagaac
     7561 atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc
     7621 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
     7681 ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc
     7741 ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc
     7801 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
     7861 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc
     7921 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac
     7981 cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc
     8041 catctgatcg caaacgccaa gaccacatat agatccaaga aacccgctaa gaacctcaag
     8101 atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag
     8161 acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg
     8221 gggcacaagc ttaattcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     8281 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     8341 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     8401 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     8461 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     8521 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     8581 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     8641 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     8701 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     8761 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     8821 cctccccgcc tggtaccttt aagaccaatg acttacaagg ca
//
