LOCUS       sequence-52983-e        8592 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 8592)
  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 8592)
  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-cmv-cfp-24xpp7
FEATURES             Location/Qualifiers
     source          1..8592
                     /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"
     enhancer        2227..2530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2531..2734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2685..2705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2731..2755
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2821..2830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2827..2829,2830..2832,2833..3546)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2872..2893)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3133..3152)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3480..3501
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4982..5570
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5035..5055)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5453..5464)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5645..5878
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5997..6015)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6083..6187
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6188..6256,6257..7048)
                     /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(6406..6425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7219..7807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7708..7727
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7961..7978
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8218..8239
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8254..8259,8260..8277,8278..8284)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8292..8308
                     /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     8297..8319
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8316..8332
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8316..8332
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8382..8401
                     /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 gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccgc accaccggtc
     2821 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2881 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2941 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     3001 cccaccctcg tgaccaccct gacctggggc gtgcagtgct tcagccgcta ccccgaccac
     3061 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3121 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3181 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3241 gggcacaagc tggagtacaa ctacatcagc cacaacgtct atatcaccgc cgacaagcag
     3301 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3361 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3421 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3481 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3541 aagtaaagcg gccggatcct aaggtaccta attgcctaga aaggagcaga cgatatggcg
     3601 tcgctccctg caggtcgact ctagaaacca gcagagcata tgggctcgct ggctgcagta
     3661 ttcccgggtt cattagatcc taaggtacct aattgcctag aaaggagcag acgatatggc
     3721 gtcgctccct gcaggtcgac tctagaaacc agcagagcat atgggctcgc tggctgcagt
     3781 attcccgggt tcattagatc ctaaggtacc taattgccta gaaaggagca gacgatatgg
     3841 cgtcgctccc tgcaggtcga ctctagaaac cagcagagca tatgggctcg ctggctgcag
     3901 tattcccggg ttcattagat cctaaggtac ctaattgcct agaaaggagc agacgatatg
     3961 gcgtcgctcc ctgcaggtcg actctagaaa ccagcagagc atatgggctc gctggctgca
     4021 gtattcccgg gttcattaga tcctaaggta cctaattgcc tagaaaggag cagacgatat
     4081 ggcgtcgctc cctgcaggtc gactctagaa accagcagag catatgggct cgctggctgc
     4141 agtattcccg ggttcattag atcctaaggt acctaattgc ctagaaagga gcagacgata
     4201 tggcgtcgct ccctgcaggt cgactctaga aaccagcaga gcatatgggc tcgctggctg
     4261 cagtattccc gggttcatta gatcctaagg tacctaattg cctagaaagg agcagacgat
     4321 atggcgtcgc tccctgcagg tcgactctag aaaccagcag agcatatggg ctcgctggct
     4381 gcagtattcc cgggttcatt agatcctaag gtacctaatt gcctagaaag gagcagacga
     4441 tatggcgtcg ctccctgcag gtcgactcta gaaaccagca gagcatatgg gctcgctggc
     4501 tgcagtattc ccgggttcat tagatcctaa ggtacctaat tgcctagaaa ggagcagacg
     4561 atatggcgtc gctccctgca ggtcgactct agaaaccagc agagcatatg ggctcgctgg
     4621 ctgcagtatt cccgggttca ttagatccta aggtacctaa ttgcctagaa aggagcagac
     4681 gatatggcgt cgctccctgc aggtcgactc tagaaaccag cagagcatat gggctcgctg
     4741 gctgcagtat tcccgggttc attagatcct aaggtaccta attgcctaga aaggagcaga
     4801 cgatatggcg tcgctccctg caggtcgact ctagaaacca gcagagcata tgggctcgct
     4861 ggctgcagta ttcccgggtt cattagatcc taaggtacct aattgcctag aaaggagcag
     4921 acgatatggc gtcgctccct gcaggtcgac tctagaaacc agcagagcat atgggctcgc
     4981 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5041 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5101 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5161 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5221 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5281 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5341 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5401 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5461 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5521 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgagatcct
     5581 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     5641 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     5701 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     5761 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     5821 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     5881 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     5941 agtgagaggc ccgggttaat taaggaaagg gctagatcat tcttgaagac gaaagggcct
     6001 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6061 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6121 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6181 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6241 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6301 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6361 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6421 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6481 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6541 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6601 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6661 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6721 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6781 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6841 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6901 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6961 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7021 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7081 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7141 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7201 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7261 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7321 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7381 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7441 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7501 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7561 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7621 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7681 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7741 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7801 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7861 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     7921 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     7981 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     8041 cagcaagctc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     8101 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     8161 ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     8221 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     8281 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac
     8341 gaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     8401 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
     8461 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
     8521 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
     8581 aaacttagta gt
//
