LOCUS       sequence-262076-        8422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ATP measurement, nucleus, lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8422)
  AUTHORS   Yaginuma H,  Okada Y
  TITLE     Live cell imaging of metabolic heterogeneity by quantitative 
            fluorescent ATP indicator protein, QUEEN-37C
  JOURNAL   bioRxiv 2021.10.08.463131
REFERENCE   2  (bases 1 to 8422)
  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: "bioRxiv 
            2021.10.08.463131"
COMMENT     Sequence Label: pLBS.EF NLS-QUE37C
FEATURES             Location/Qualifiers
     source          1..8422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1802..1918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1972..3154
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2203..3145
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3102..3122
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     3163..3179
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3164..3180
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3210..3230
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3243..3252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3609..3860
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     3797..3818
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3924..3945)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4185..4204)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4431..5019
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4484..4504)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4902..4913)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5110..5343
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5415..5496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5452..5471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5506..5525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(5574..5592)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5613..5629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5613..5629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5626..5648)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5637..5653
                     /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(5661..5667,5668..5685,5686..5691))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5706..5727
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5844..5861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6015..6603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6095..6114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6774..7565,7566..7634))
                     /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     7397..7416
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7635..7739)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7765..8220)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7902..7923)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8114..8133
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8347..8369
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8361..8378
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8362..8378
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8385..8404
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8385..8403
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttgg
     1921 gcccttccaa ccttccaacc gggattgatt attgactaga tcatcgcgtg aggctccggt
     1981 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     2041 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2101 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2161 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     2221 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacg
     2281 cccctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
     2341 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
     2401 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
     2461 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
     2521 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
     2581 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
     2641 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
     2701 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     2761 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     2821 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     2881 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     2941 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     3001 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     3061 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     3121 tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt agtcgaggtc gacggtatcg
     3181 ataagcttga tatcgaattc accatggcgc cgaaaaaaaa acgcaaagtg gatccaccgg
     3241 tcgccaccat ggtgaaaact gtgaaagtga atataacaac ccctgatggg ccagtctacg
     3301 acgctgatat cgagatggtg tccgtgcggg ccgagagtgg tgatctcggc atcctccccg
     3361 gtcacattcc cacaaaggcc ccactgaaga tcggagctgt gcggctgaag aaggacggcc
     3421 aaacagagga tgtcgcagtc tcaggcggca ctgttgaagt gcggcctgac cacgttacca
     3481 ttaatgctca agccgctgaa cgggcggagg acattgacgt cctccgcgcc aaagcggcga
     3541 aagagcgggc ggagcgccgc ctgcaaagcc agacccgtta caacagccac aacgtctata
     3601 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     3661 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     3721 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     3781 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3841 gcatggacga gctgtacaag ggtggcaccg gagggagcat ggtgagcaag ggcgaggagc
     3901 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3961 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     4021 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     4081 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     4141 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     4201 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     4261 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacctgggtg
     4321 acatcgactt caaacgggcc gaactggcgt taaaacgcgc catgaaccgt ttgagcgttg
     4381 cggaagataa gtgagcggcc gcgactctag atcactagtc tcgactcgac aatcaacctc
     4441 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4501 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4561 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4621 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4681 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4741 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4801 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     4861 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4921 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcatctt cgccttcgcc
     4981 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat tcgagctcgg
     5041 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     5101 aggggggact ggaagggcta attcactccc aacgaagata agatctgctt tttgcttgta
     5161 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5221 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5281 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5341 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     5401 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5521 tcatcaatgt atctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     5581 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5641 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5701 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5761 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5821 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5881 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5941 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6541 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6601 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6661 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6721 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6781 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6841 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6901 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6961 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7021 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7081 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7141 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7201 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7261 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7321 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7381 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7441 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7501 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7561 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7621 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7681 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7741 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     7801 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     7861 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     7921 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     7981 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     8041 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     8101 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     8161 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     8221 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     8281 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     8341 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat
     8401 agggcgaatt gggtaccacg cg
//
