LOCUS       sequence-244580-        5753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses transmembrane intracellular NanoLuc in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5753)
  AUTHORS   Kim CK, Cho KF, Kim MW, Ting AY
  TITLE     Luciferase-LOV BRET enables versatile and specific transcriptional 
            readout of cellular protein-protein interactions.
  JOURNAL   eLife. 2019 Apr 3;8. pii: 43826. doi: 10.7554/eLife.43826.
  PUBMED    30942168
REFERENCE   2  (bases 1 to 5753)
  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: "2019-04-3"; volume: "8. pii: 43826. doi"; pages: " 
            10.7554/eLife.43826"
COMMENT     Sequence Label: pAAV-IgK sp-HA-CD4-10aa linker-CIBN-NNES-NanoLuc
FEATURES             Location/Qualifiers
     source          1..5753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        505..708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     659..679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     753..772
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        753..771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      791..800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     797..856
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             881..907
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             908..934
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1682..2191
                     /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             2231..2743
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2774..2791)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2780..2987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3016..3156
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3016..3145
                     /label=AAV2 ITR
     rep_origin      3231..3686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3318..3337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3528..3549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3703..3722)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3822..3844
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3882..3900)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3968..4072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4073..4141,4142..4933)
                     /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     complement(4291..4310)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5104..5692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5593..5612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      781 gcttgcggcc gccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
      841 tccaggttcc actggtgacg gggcccagcc ggccagatct tacccatacg atgtgccaga
      901 ttacgcctat ccctatgacg tccctgatta cgctctcgac ttccagaagg cctccagcat
      961 agtctataag aaagaggggg aacaggtgga gttctccttc ccactcgcct ttacagttga
     1021 aaagctgacg ggcagtggcg agctgtggtg gcaggcggag agggcttcct cctccaagtc
     1081 ttggatcacc tttgacctga agaacaagga agtgtctgta aaacgggtta cccaggaccc
     1141 taagctccag atgggcaaga agctcccgct ccacctcacc ctgccccagg ccttgcctca
     1201 gtatgctggc tctggaaacc tcaccctggc ccttgaagcg aaaacaggaa agttgcatca
     1261 ggaagtgaac ctggtggtga tgagagccac tcagctccag aaaaatttga cctgtgaggt
     1321 gtggggaccc acctccccta agctgatgct gagcttgaaa ctggagaaca aggaggcaaa
     1381 ggtctcgaag cgggagaagg cggtgtgggt gctgaaccct gaggcgggga tgtggcagtg
     1441 tctgctgagt gactcgggac aggtcctgct ggaatccaac atcaaggttc tgcccacatg
     1501 gtccaccccg gtgcagccaa tggccctgat tgtgctgggg ggcgtcgccg gcctcctgct
     1561 tttcattggg ctaggcatct tcttctgtgt caggtgccgg caccgaaggc gcaagggctc
     1621 gggctcgacc tcgggctcgg gcagcggtgg ctctagaggt tctggtggca gctctggagg
     1681 tatgaatgga gctataggag gtgacctttt gctcaatttt cctgacatgt cggtcctaga
     1741 gcgccaaagg gctcacctca agtacctcaa tcccaccttt gattctcctc tcgccggctt
     1801 ctttgccgat tcttcaatga ttaccggcgg cgagatggac agctatcttt cgactgccgg
     1861 tttgaatctt ccgatgatgt acggtgagac gacggtggaa ggtgattcaa gactctcaat
     1921 ttcgccggaa acgacgcttg ggactggaaa tttcaaggca gcgaagtttg atacagagac
     1981 taaggattgt aatgaggcgg cgaagaagat gacgatgaac agagatgacc tagtagaaga
     2041 aggagaagaa gagaagtcga aaataacaga gcaaaacaat gggagcacaa aaagcatcaa
     2101 gaagatgaaa cacaaagcca agaaagaaga gaacaatttc tctaatgatt catctaaagt
     2161 gacgaaggaa ttggagaaaa cggattatat tcatgagctg gccgagaagc tggccggcct
     2221 ggacatcaac gtcttcacac tcgaagattt cgttggggac tggcgacaga cagccggcta
     2281 caacctggac caagtccttg aacagggagg tgtgtccagt ttgtttcaga atctcggggt
     2341 gtccgtaact ccgatccaaa ggattgtcct gagcggtgaa aatgggctga agatcgacat
     2401 ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat
     2461 ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc actatggcac
     2521 actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc cgtatgaagg
     2581 catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga acggcaacaa
     2641 aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag taaccatcaa
     2701 cggagtgacc ggctggcggc tgtgcgaacg cattctggcg tgagaattcc tcgagctaga
     2761 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2821 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2881 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2941 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggaggt aaccacgtgc
     3001 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3061 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3121 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     3181 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     3241 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3301 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3361 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3421 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3481 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3541 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3601 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3661 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3721 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3781 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3841 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3901 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3961 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4021 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4081 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4141 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4201 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4261 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4321 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4381 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4441 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4501 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4561 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4621 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4681 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4741 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat
     4801 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4861 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4921 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4981 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5041 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5101 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5161 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5221 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5281 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5341 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5401 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5461 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5521 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5581 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5641 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5701 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
