LOCUS       sequence-199089-        5627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector, 25 repeats of 19 bp BoxB sequence from 
            bacteriophage-lambda, for RNA labeling. BoxB-RNA is bound by 22 
            amino acid RNA binding domain of lambda-anti-terminator protein N, 
            e.g. tagged with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5627)
  AUTHORS   Martin RM, Rino J, Carvalho C, Kirchhausen T, Carmo-Fonseca M
  TITLE     Live-cell visualization of pre-mRNA splicing with single-molecule 
            sensitivity.
  JOURNAL   Cell Rep. 2013 Sep 26;4(6):1144-55. doi: 
            10.1016/j.celrep.2013.08.013. Epub 2013 Sep 12.
  PUBMED    24035393
REFERENCE   2  (bases 1 to 5627)
  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"; doi: 
            "10.1016/j.celrep.2013.08.013"; journalName: "Cell Rep"; date: 
            "2013-09-26- 26"; volume: "4"; issue: "6"; pages: "1144-55"
COMMENT     Sequence Label: pCMV5-25BoxB
FEATURES             Location/Qualifiers
     source          1..5627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        178..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    805..823
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    849..867
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    873..891
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    932..950
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    956..974
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1012..1030
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1056..1074
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1080..1098
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1139..1157
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1163..1181
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1219..1237
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1263..1281
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1287..1305
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1346..1364
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1370..1388
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1426..1444
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1495..1513
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1554..1572
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1578..1596
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1634..1652
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1678..1696
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1702..1720
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1761..1779
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1785..1803
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     polyA_signal    1828..2450
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2027..2046)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2474..2494)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2479..2808
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2659..2794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2846..2865)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2847..2865)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2879..2895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2879..2895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2892..2914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2903..2919
                     /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(2927..2933,2934..2951,2952..2957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2972..2993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3110..3127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3281..3869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3361..3380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4040..4831,4832..4900))
                     /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     4663..4682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4901..5005)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5032..5487
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5119..5138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5329..5350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(5613..5627,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5627,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
       61 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
      121 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
      181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      781 attcagatct aactggggat tcctgggccc tgaagaaggg cccctcgact aagtccaact
      841 actaaactgg gccctgaaga agggcccata tagggccctg aagaagggcc ctatcgagga
      901 tattatctcg actaagtcca actactaaac tgggccctga agaagggccc atatagggcc
      961 ctgaagaagg gccctatcga ggatattatc tggatctaac tggggattcc tgggccctga
     1021 agaagggccc ctcgactaag tccaactact aaactgggcc ctgaagaagg gcccatatag
     1081 ggccctgaag aagggcccta tcgaggatat tatctcgact aagtccaact actaaactgg
     1141 gccctgaaga agggcccata tagggccctg aagaagggcc ctatcgagga tattatctgg
     1201 atctaactgg ggattcctgg gccctgaaga agggcccctc gactaagtcc aactactaaa
     1261 ctgggccctg aagaagggcc catatagggc cctgaagaag ggccctatcg aggatattat
     1321 ctcgactaag tccaactact aaactgggcc ctgaagaagg gcccatatag ggccctgaag
     1381 aagggcccta tcgaggatat tatctggatc taactgggga ttcctgggcc ctgaagaagg
     1441 gcccctcgac taagtccaac tactaaactg ggccctgaaa gaagggccca tatagggccc
     1501 tgaagaaggg ccctatcgag gatattatct cgactaagtc caactactaa actgggccct
     1561 gaagaagggc ccatataggg ccctgaagaa gggccctatc gaggatatta tctggatcta
     1621 actggggatt cctgggccct gaagaagggc ccctcgacta agtccaacta ctaaactggg
     1681 ccctgaagaa gggcccatat agggccctga agaagggccc tatcgaggat attatctcga
     1741 ctaagtccaa ctactaaact gggccctgaa gaagggccca tatagggccc tgaagaaggg
     1801 ccctatcgag gatattatct ggatcccggg tggcatccct gtgacccctc cccagtgcct
     1861 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     1921 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     1981 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     2041 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     2101 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     2161 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     2221 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     2281 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     2341 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     2401 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     2461 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2521 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2641 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2701 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2761 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcctcgag
     2821 gaactgaaaa accagaaagt taattcccta tagtgagtcg tattaaattc gtaatcatgt
     2881 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2941 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3001 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3061 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3721 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     5041 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5101 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5161 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5221 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5281 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5341 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5401 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5461 ttaacaaaat attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga
     5521 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5581 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
