LOCUS       sequence-415908-        6099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CIB1 fused with Split-TurboID-N and pmGFP in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  AUTHORS   Shafraz O, Davis CMO, Sivasankar S
  TITLE     Light-activated BioID - an optically activated proximity labeling 
            system to study protein-protein interactions.
  JOURNAL   J Cell Sci. 2023 Oct 1;136(19):jcs261430. doi: 10.1242/jcs.261430. 
            Epub 2023 Oct 11.
  PUBMED    37756605
REFERENCE   2  (bases 1 to 6099)
  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: "J Cell 
            Sci."; date: "2023-10-1"; volume: "136(19):jcs261430. doi"; pages: "
            10.1242/jcs.261430. Epub 2023 Oct 11"
COMMENT     Sequence Label: CibN
FEATURES             Location/Qualifiers
     source          1..6099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..328
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        329..532
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     483..503
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             563..1072
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             1574..1615
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      1883..1892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1889..1891,1892..1894,1895..2605)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1889..1891,1892..1894,1895..2605)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1934..1955)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2195..2214)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2542..2563
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2662..2727
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2851..2972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2888..2907)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2942..2961
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2979..3434)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3116..3137)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3328..3347
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3461..3565
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3567..3924
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3567..3758
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3590..3610)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3775..3910
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3959..4753
                     /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             3959..4753
                     /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(4013..4032)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4623..4642
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4941..4960)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4985..5032
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5361..5949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5850..5869
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
       61 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      121 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      181 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      241 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      301 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      361 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      421 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      481 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      541 gaaccgtcag atccgctagc ccatgaatgg agctatagga ggtgaccttt tgctcaattt
      601 tcctgacatg tcggtcctag agcgccaaag ggctcacctc aagtacctca atcccacctt
      661 tgattctcct ctcgccggct tctttgccga ttcttcaatg attaccggcg gcgagatgga
      721 cagctatctt tcgactgccg gtttgaatct tccgatgatg tacggtgaga cgacggtgga
      781 aggtgattca agactctcaa tttcgccgga aacgacgctt gggactggaa atttcaaggc
      841 agcgaagttt gatacagaga ctaaggattg taatgaggcg gcgaagaaga tgacgatgaa
      901 cagagatgac ctagtagaag aaggagaaga agagaagtcg aaaataacag agcaaaacaa
      961 tgggagcaca aaaagcatca agaagatgaa acacaaagcc aagaaagaag agaacaattt
     1021 ctctaatgat tcatctaaag tgacgaagga attggagaaa acggattata ttcatgttcg
     1081 tgcacgacga ggccaagcca ctgatagtca cagcatagca gaacgagtta gaagagaaaa
     1141 gatcagtgag agaatgaagt ttctacaaga tttggttcct ggatgcgaca agatcacagg
     1201 caaagcaggg atgcttgatg aaatcattaa ctatgttcag tctcttcaga gacaaatcga
     1261 gttcttatcg atgaaactag caattgtgaa tccaaggccg gattttgata tggatgacat
     1321 ttttgccaaa gaggttgcct caactccaat gactgtggtg ccatctcctg aaatggttct
     1381 ttccggttat tctcatgaga tggttcactc tggttattct agtgagatgg ttaactccgg
     1441 ttaccttcat gtcaatccaa tgcagcaagt gaataccagt tctgatccat tgtcatgctt
     1501 caacaatggc gaagctcctt cgatgtggga ctctcatgtg cagaatctct atggcaattt
     1561 aggagtagat atcggcaagc ccatccccaa ccccctgctg ggcctggaca gcaccaaggg
     1621 ctcgggctcg acctcgggct cgggcaccgg taaagacaat actgtgcctc tgaagctgat
     1681 cgctctcctg gctaatggcg agttccatag tggcgaacag ctgggagaaa ccctgggcat
     1741 gtccagggcc gctatcaaca agcacattca gactctgcgc gactggggcg tggacgtgtt
     1801 caccgtgccc ggaaagggct actctctgcc cgagcctatc ccgctgctga acgctaaaca
     1861 gattctgggg agtggaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1921 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1981 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2041 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2101 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2161 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2221 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2281 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2341 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2401 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2461 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2521 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2581 tctcggcatg gacgagctgt ataagggtaa aaagaagaaa aagaagtcaa agacaaagtg
     2641 tgtaattatg tagtgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca
     2701 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     2761 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2821 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2881 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2941 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3001 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3061 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3121 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3181 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3241 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3301 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3361 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3421 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3481 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3541 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3601 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3661 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3721 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3781 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3841 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3901 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     3961 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4021 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4081 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4141 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4201 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4261 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4321 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4381 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4441 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4501 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4561 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4621 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4681 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4741 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4801 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4861 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4921 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4981 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5041 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5101 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5161 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5221 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5281 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5341 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5401 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5461 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5521 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5581 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5641 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5701 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5761 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5821 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5881 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5941 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6001 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6061 cattagttat taatagtaat caattacggg gtcattagt
//
