LOCUS       sequence-384648-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mAzurite mRNA with 16xRhoBAST in 3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5797)
  AUTHORS   Englert D, Burger EM, Grun F, Verma MS, Lackner J, Lampe M, Buhler 
            B, Schokolowski J, Nienhaus GU, Jaschke A, Sunbul M
  TITLE     Fast-exchanging spirocyclic rhodamine probes for aptamer-based 
            super-resolution RNA imaging.
  JOURNAL   Nat Commun. 2023 Jun 30;14(1):3879. doi: 10.1038/s41467-023-39611-1.
  PUBMED    37391423
REFERENCE   2  (bases 1 to 5797)
  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: "Nat 
            Commun."; date: "2023-06-30"; volume: "14(1):3879. doi"; pages: " 
            10.1038/s41467-023-39611-1"
COMMENT     Sequence Label: mAzurite-16xRhoBAST
FEATURES             Location/Qualifiers
     source          1..5797
                     /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..1404)
                     /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    2657..2778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2694..2713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2748..2767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2785..3240)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2922..2943)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3134..3153
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3267..3371
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3373..3730
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3373..3564
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3396..3416)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3581..3716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3765..4559
                     /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             3765..4559
                     /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(3819..3838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4429..4448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4747..4766)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4791..4838
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5167..5755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5656..5675
                     /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 gtaaggactc agatctcgag ctcaagcttc gaattctgca
     1441 gtcgacatag aggaacctcc gcgaaagcgg tgaaggagag gcgcaaggtt aaccgcctca
     1501 ggttcctcat aacaaggcct ccgcgaaagc ggtgaaggag cggcacaagg ttaactgccg
     1561 caggccttgt atactcgaca taggaagacc ttcgcgaaag cgatgaagga gcggtgcaag
     1621 gttaaccacc gcaggtcttc cataagcaga ccttcgcgaa agcgatgaag gagtggcgca
     1681 aggttaaccg ccacaggtct gctatactcg acatagagga acctccgcga aagcggtgaa
     1741 ggagaggcgc aaggttaacc gcctcaggtt cctcataaca aggcctccgc gaaagcggtg
     1801 aaggagcggc acaaggttaa ctgccgcagg ccttgtatac tcgacatagg aagaccttcg
     1861 cgaaagcgat gaaggagcgg tgcaaggtta accaccgcag gtcttccata agcagacctt
     1921 cgcgaaagcg atgaaggagt ggcgcaaggt taaccgccac aggtctgcta tactcgacat
     1981 agaggaacct ccgcgaaagc ggtgaaggag aggcgcaagg ttaaccgcct caggttcctc
     2041 ataacaaggc ctccgcgaaa gcggtgaagg agcggcacaa ggttaactgc cgcaggcctt
     2101 gtatactcga cataggaaga ccttcgcgaa agcgatgaag gagcggtgca aggttaacca
     2161 ccgcaggtct tccataagca gaccttcgcg aaagcgatga aggagtggcg caaggttaac
     2221 cgccacaggt ctgctatact cgacatagag gaacctccgc gaaagcggtg aaggagaggc
     2281 gcaaggttaa ccgcctcagg ttcctcataa caaggcctcc gcgaaagcgg tgaaggagcg
     2341 gcacaaggtt aactgccgca ggccttgtat actcgacata ggaagacctt cgcgaaagcg
     2401 atgaaggagc ggtgcaaggt taaccaccgc aggtcttcca taagcagacc ttcgcgaaag
     2461 cgatgaagga gtggcgcaag gttaaccgcc acaggtctgc tatactcgac gcgtgaattc
     2521 actcctcgga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     2581 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2641 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2701 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2761 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2821 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2881 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2941 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3001 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3061 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3121 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3181 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3241 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3301 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3361 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3481 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3541 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3601 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3661 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3721 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3781 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3841 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3901 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3961 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4021 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4081 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4141 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4201 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4261 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4321 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4381 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4441 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4501 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4561 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4621 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4681 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4741 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4801 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4861 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4921 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4981 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5041 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5101 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5161 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5221 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5281 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5341 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5401 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5461 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5521 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5581 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5641 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5701 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5761 cagcaacgcg gcctttttac ggttcctggc cttttgc
//
