LOCUS       sequence-384646-        7790 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of mAzurite mRNA with 8xbiRhoBAST-24xMS2v6 in 3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7790)
  AUTHORS   Buhler B, Schokolowski J, Benderoth A, Englert D, Grun F, Jaschke A,
            Sunbul M
  TITLE     Avidity-based bright and photostable light-up aptamers for 
            single-molecule mRNA imaging.
  JOURNAL   Nat Chem Biol. 2023 Apr;19(4):478-487. doi: 
            10.1038/s41589-022-01228-8. Epub 2023 Jan 19.
  PUBMED    36658339
REFERENCE   2  (bases 1 to 7790)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41589-022-01228-8"; journalName: "Nat Chem Biol"; date: 
            "2023-04"; volume: "19"; issue: "4"; pages: "478-487"
COMMENT     Sequence Label: mAzurite-8xbiRhoBAST-24xMS2v6
FEATURES             Location/Qualifiers
     source          1..7790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        133..436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        437..640
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     591..611
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(685..687,688..690,691..1401)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNIYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFRTAAGITHGMDELYK"
     primer_bind     complement(730..751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(991..1010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    4650..4771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4687..4706)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4741..4760
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4778..5233)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4915..4936)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5127..5146
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5260..5364
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5366..5723
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5366..5557
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5389..5409)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5574..5709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5758..6552
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5758..6552
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5812..5831)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6422..6441
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6740..6759)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6784..6831
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7160..7748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7649..7668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
       61 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      121 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      181 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      241 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      301 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      361 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      421 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      481 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      541 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      601 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      661 ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      841 aagctgcccg tgccctggcc caccctcgtg accaccctga gccacggcgt gcagtgcttc
      901 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      961 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1081 gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca caacatctat
     1141 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1201 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1261 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     1321 aacgagaagc gcgatcacat ggtcctgctg gagttccgca ccgccgccgg gatcactcac
     1381 ggcatggacg agctgtacaa gtccggactc agatctcgac gggtctagag gtatgtggga
     1441 ggaacctccg cgaaagcggt gaaggagagg cgcaaggtta accgcctcag gttccaggag
     1501 gattcgtcct ccaaaggcct ccgcgaaagc ggtgaaggag cgccacaagg ttaactggcg
     1561 caggccttac ccacataccg ctagaggagc ccggaaggaa cctccgcgaa agcggtgaag
     1621 gagaggcgca aggttaaccg cctcaggttc cagcggagtt cgctccgcaa aggcctccgc
     1681 gaaagcggtg aaggagcgcc acaaggttaa ctggcgcagg ccttatccgg gctccgctag
     1741 aggtatgtgg gaggaacctc cgcgaaagcg gtgaaggaga ggcgcaaggt taaccgcctc
     1801 aggttccagg aggattcgtc ctccaaaggc ctccgcgaaa gcggtgaagg agcgccacaa
     1861 ggttaactgg cgcaggcctt acccacatac cgctagagga gcccggaagg aacctccgcg
     1921 aaagcggtga aggagaggcg caaggttaac cgcctcaggt tccagcggag ttcgctccgc
     1981 aaaggcctcc gcgaaagcgg tgaaggagcg ccacaaggtt aactggcgca ggccttatcc
     2041 gggctccgct agaggtatgt gggaggaacc tccgcgaaag cggtgaagga gaggcgcaag
     2101 gttaaccgcc tcaggttcca ggaggattcg tcctccaaag gcctccgcga aagcggtgaa
     2161 ggagcgccac aaggttaact ggcgcaggcc ttacccacat accgctagag gagcccggaa
     2221 ggaacctccg cgaaagcggt gaaggagagg cgcaaggtta accgcctcag gttccagcgg
     2281 agttcgctcc gcaaaggcct ccgcgaaagc ggtgaaggag cgccacaagg ttaactggcg
     2341 caggccttat ccgggctccg ctagaggtat gtgggaggaa cctccgcgaa agcggtgaag
     2401 gagaggcgca aggttaaccg cctcaggttc caggaggatt cgtcctccaa aggcctccgc
     2461 gaaagcggtg aaggagcgcc acaaggttaa ctggcgcagg ccttacccac ataccgctag
     2521 aggagcccgg aaggaacctc cgcgaaagcg gtgaaggaga ggcgcaaggt taaccgcctc
     2581 aggttccagc ggagttcgct ccgcaaaggc ctccgcgaaa gcggtgaagg agcgccacaa
     2641 ggttaactgg cgcaggcctt atccgggctc cgctagagcg gacttcggtc cgctttttac
     2701 taggacctgc aggcatgcaa gcttgacgtc ggttaccgat atccatatgg cggccgcatc
     2761 gatctcgagc tcaagcttcg aattctgcag tcgacgggcc cgggatccga tccaatcatg
     2821 gatcccagag ccccctggca atcgcgggga gacgcaggac taccgcgtct tcactctcgc
     2881 ttgcgcggta tacgcaggga gcaacgtcac ccagggcgac acagaggaat accctgtgtg
     2941 gccccacggt gctacaaaga acattccgta ttgctcacca gtgtcacatg tggaggacta
     3001 ccccacaagc gagtcaggaa accttcgggg catcgcacca ttatcccgaa caatcgacac
     3061 gcaggattac cgcgtgggac actctgttcc ccgtcaaaat tggaccatac cggagtcggc
     3121 ttacgtcatg caggattacc gcatgcatgg tgcagaatat cggcatgtct actcgtacag
     3181 tcaaatctac tcgtgtggtc aggactaccg accacttcta ttctattcat cttttcggtt
     3241 gtgcaggtat tctgccgatg tacgagaaga cgattacgct tctcgactac cttcatcata
     3301 catggtgtgc agatggccgc caagtttttt gccaatggag gaatacccca ttctctgtca
     3361 accaacccat gcaaagttta cactctgcta tggcagcact gtcgcagagg aataccctgc
     3421 gagccaaaac ggcccccggt gcgtgtattg catctgcctt gcgagcattc acagggacga
     3481 atacgccctg ccgttgcatt acttcaatat gggtgctctg tcgtcgtcat caggaccatt
     3541 tgcgcaggac taccgcgcat atatcatcag cactcgtgcg gatacttctg ggattcctat
     3601 tgttacgcga gctcaggaat accgagctct ggcgacagag accctcacac ggaagatccc
     3661 agagccccct ggcaatcgcg gggagacgca ggactaccgc gtcttcactc tcgcttgcgc
     3721 ggtatacgca gggagcaacg tcacccaggg cgacacagag gaataccctg tgtggcccca
     3781 cggtgctaca aagaacattc cgtattgctc accagtgtca catgtggagg actaccccac
     3841 aagcgagtca ggaaaccttc ggggcatcgc accattatcc cgaacaatcg acacgcagga
     3901 ttaccgcgtg ggacactctg ttccccgtca aaattggacc ataccggagt cggcttacgt
     3961 catgcaggat taccgcatgc atggtgcaga atatcggcat gtctactcgt acagtcaaat
     4021 ctactcgtgt ggtcaggact accgaccact tctattctat tcatcttttc ggttgtgcag
     4081 gtattctgcc gatgtacgag aagacgatta cgcttctcga ctaccttcat catacatggt
     4141 gtgcagatgg ccgccaagtt ttttgccaat ggaggaatac cccattctct gtcaaccaac
     4201 ccatgcaaag tttacactct gctatggcag cactgtcgca gaggaatacc ctgcgagcca
     4261 aaacggcccc cggtgcgtgt attgcatctg ccttgcgagc attcacaggg acgaatacgc
     4321 cctgccgttg cattacttca atatgggtgc tctgtcgtcg tcatcaggac catttgcgca
     4381 ggactaccgc gcatatatca tcagcactcg tgcggatact tctgggattc ctattgttac
     4441 gcgagctcag gaataccgag ctctggcgac agagaccctc acacggaaga tctatcgatc
     4501 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     4561 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4621 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4681 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4741 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     4801 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4861 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4921 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4981 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5041 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5101 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5161 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5221 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5281 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5341 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     5401 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5461 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5521 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5581 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5641 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5701 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     5761 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5821 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5881 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     5941 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6001 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6061 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6121 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6181 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6241 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     6301 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6361 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6421 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6481 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6541 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6601 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6661 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6721 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     6781 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     6841 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     6901 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     6961 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     7021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
//
