LOCUS       sequence-260988-        8163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cytosol marker, lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8163)
  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 8163)
  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-iRFP
FEATURES             Location/Qualifiers
     source          1..8163
                     /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"
     misc_feature    4172..4760
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4225..4245)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4643..4654)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4851..5084
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5156..5237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5193..5212)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5247..5266
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(5315..5333)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5354..5370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5354..5370)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5367..5389)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5378..5394
                     /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(5402..5408,5409..5426,5427..5432))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5447..5468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5585..5602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5756..6344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5836..5855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6515..7306,7307..7375))
                     /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     7138..7157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7376..7480)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7506..7961)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7643..7664)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7855..7874
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8088..8110
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8102..8119
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8103..8119
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8126..8145
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8126..8144
                     /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 atggccgaag ggagcgtggc aagacagccc gatctgctga
     3241 catgcgacga cgaaccaatc cacattccag gagctatcca gcctcatgga ctgctgctgg
     3301 ctctggccgc tgacatgact atcgtggcag gctctgataa cctgccagag ctgaccgggc
     3361 tggctatcgg agcactgatt ggccggtccg cagccgacgt gttcgattct gagactcaca
     3421 atagactgac catcgccctg gctgaacctg gggctgcagt gggagcccca attaccgtcg
     3481 gcttcacaat gcgaaaggac gctggcttta tcgggtcctg gcaccgccat gatcagctga
     3541 tttttctgga gctggaacca cctcagagag acgtggcaga gccacaggct ttctttcgga
     3601 gaacaaacag cgccatcagg cgcctgcagg ccgctgagac tctggaatcc gcttgcgcag
     3661 ccgctgcaca ggaagtgagg aagatcaccg ggttcgatcg cgtcatgatc tacagattcg
     3721 cctcagactt tagcggagaa gtgatcgcag aagatcgctg tgccgaggtc gaaagtaaac
     3781 tgggcctgca ttacccagcc tcaaccgtgc ctgcacaggc acgacggctg tatacaatca
     3841 accccgtgcg gatcattccc gacattaatt acagaccagt gcccgtcaca cctgacctga
     3901 acccagtgac tgggaggccc atcgatctga gcttcgccat tctgcgcagc gtgtcccctg
     3961 tccacctgga gtttatgagg aatatcggaa tgcatggcac catgtctatc agtattctga
     4021 gaggcgaaag gctgtggggg ctgatcgtgt gccaccatcg gacaccttac tatgtggacc
     4081 tggacggcag gcaggcttgt gaactggtgg ctcaggtgct ggcttggcag attggcgtga
     4141 tgaaggagga ataaactagt ctcgactcga caatcaacct ctggattaca aaatttgtga
     4201 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     4261 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4321 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4381 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     4441 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     4501 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4561 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4621 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4681 gctgccggct ctgcggcctc ttccgcatct tcgccttcgc cctcagacga gtcggatctc
     4741 cctttgggcc gcctccccgc ctggaattaa ttcgagctcg gtacctttaa gaccaatgac
     4801 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     4861 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga
     4921 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     4981 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     5041 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     5101 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     5161 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5221 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatctagagc
     5281 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta atttcgagct
     5341 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5401 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5461 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5521 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6181 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6361 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6781 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7141 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7201 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7261 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7321 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7381 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7441 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7501 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     7561 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     7621 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     7681 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     7741 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     7801 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     7861 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     7921 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     7981 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     8041 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     8101 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccac
     8161 gcg
//
