LOCUS       sequence-52979-e        7805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7805)
  AUTHORS   Wu B, Chao JA, Singer RH
  TITLE     Fluorescence fluctuation spectroscopy enables quantitative imaging 
            of single mRNAs in living cells.
  JOURNAL   Biophys J. 2012 Jun 20;102(12):2936-44. Epub 2012 Jun 19.
  PUBMED    22735544
REFERENCE   2  (bases 1 to 7805)
  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: "Biophys 
            J."; date: "2012-06-20"; volume: "102(12)"; pages: "2936-44. Epub 
            2012 Jun 19"
COMMENT     Sequence Label: phage-ubc-nls-ha-tdPCP-gfp
FEATURES             Location/Qualifiers
     source          1..7805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2218..2616
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      2619..2628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2631..2651
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2655..2681
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2655..2681
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2682..2693
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3456..3465
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3462..3464,3465..3467,3468..4181)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3462..3464,3465..3467,3468..4178)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3507..3528)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3768..3787)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4115..4136
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4195..4783
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4248..4268)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4666..4677)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4858..5091
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5210..5228)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5296..5400
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5401..5469,5470..6261)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5619..5638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6432..7020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6921..6940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7174..7191
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7431..7452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7467..7472,7473..7490,7491..7497)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7505..7521
                     /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)."
     primer_bind     7510..7532
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7529..7545
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7529..7545
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7595..7614
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtggc
     2221 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca
     2281 cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     2341 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     2401 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     2461 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgccatgggc ccaaaaaaga
     2641 aaagaaaagt tggctacccc tacgacgtgc ccgactacgc catcgaaggc cgccatatgc
     2701 tagcctccaa aaccatcgtt ctttcggtcg gcgaggctac tcgcactctg actgagatcc
     2761 agtccaccgc agaccgtcag atcttcgaag agaaggtcgg gcctctggtg ggtcggctgc
     2821 gcctcacggc ttcgctccgt caaaacggag ccaagaccgc gtatcgcgtc aacctaaaac
     2881 tggatcaggc ggacgtcgtt gattccggac ttccgaaagt gcgctacact caggtatggt
     2941 cgcacgacgt gacaatcgtt gcgaatagca ccgaggcctc gcgcaaatcg ttgtacgatt
     3001 tgaccaagtc cctcgtcgcg acctcgcagg tcgaagatct tgtcgtcaac cttgtgccgc
     3061 tgggccgtgc ggatccgcta gcctccaaaa ccatcgttct ttcggtcggc gaggctactc
     3121 gcactctgac tgagatccag tccaccgcag accgtcagat cttcgaagag aaggtcgggc
     3181 ctctggtggg tcggctgcgc ctcacggctt cgctccgtca aaacggagcc aagaccgcgt
     3241 atcgcgtcaa cctaaaactg gatcaggcgg acgtcgttga ttccggactt ccgaaagtgc
     3301 gctacactca ggtatggtcg cacgacgtga caatcgttgc gaatagcacc gaggcctcgc
     3361 gcaaatcgtt gtacgatttg accaagtccc tcgtcgcgac ctcgcaggtc gaagatcttg
     3421 tcgtcaacct tgtgccgctg ggccgtccac cggtcgccac catggtgagc aagggcgagg
     3481 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3541 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3601 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3661 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3721 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3781 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3841 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3901 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     3961 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4021 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4081 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4141 ccgccgggat cactctcggc atggacgagc tgtacaagta aatcgataga tcctaatcaa
     4201 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4261 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4321 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4381 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4441 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4501 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4561 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4621 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4681 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4741 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat cctttaagac
     4801 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     4861 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     4921 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4981 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5041 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     5101 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     5161 ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata
     5221 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5281 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5341 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5401 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5461 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5521 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5581 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5641 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5701 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5761 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5821 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5881 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5941 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6001 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6061 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6121 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6181 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6241 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6301 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6361 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6541 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7081 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7141 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7201 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag
     7261 ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     7321 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg
     7381 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7441 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7501 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc
     7561 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7621 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
     7681 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
     7741 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
     7801 gtagt
//
