LOCUS       sequence-273264-        7227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7227)
  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 7227)
  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
FEATURES             Location/Qualifiers
     source          1..7227
                     /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    3236..3824
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3289..3309)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3707..3718)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3915..4148
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4220..4301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4257..4276)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4311..4330
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(4379..4397)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4418..4434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4418..4434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4431..4453)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4442..4458
                     /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(4466..4472,4473..4490,4491..4496))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4511..4532
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4649..4666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4820..5408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4900..4919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5579..6370,6371..6439))
                     /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     6202..6221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6440..6544)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6570..7025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6707..6728)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6919..6938
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7152..7174
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7166..7183
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7167..7183
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7190..7209
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7190..7208
                     /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 ctgcagcccg ggggatccac tagtctcgac tcgacaatca
     3241 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3301 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3361 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3421 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3481 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3541 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3601 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3661 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3721 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc atcttcgcct
     3781 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattcgag
     3841 ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     3901 agaaaagggg ggactggaag ggctaattca ctcccaacga agataagatc tgctttttgc
     3961 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     4021 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     4081 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     4141 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     4201 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     4261 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4321 caaactcatc aatgtatcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
     4381 tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4441 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4501 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4561 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4621 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4681 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4741 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4801 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4861 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4921 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4981 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5041 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5101 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5161 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5221 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5281 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5341 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5521 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5641 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5701 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5761 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5821 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5881 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5941 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6001 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6061 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6121 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6181 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6241 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6301 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6361 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6421 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6541 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     6601 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6661 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6721 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6781 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6841 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6901 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6961 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     7021 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     7081 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     7141 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc
     7201 actatagggc gaattgggta ccacgcg
//
