LOCUS       sequence-384631-        6268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of circular biRhoBAST .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6268)
  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 6268)
  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: 
            "10.1038/s41589-022-01228-8"; journalName: "Nat Chem Biol"; date: 
            "2023-04"; volume: "19"; issue: "4"; pages: "478-487"
COMMENT     Sequence Label: pAV-U6+27-Tornado-biRhoBAST
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..273
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     25..45
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     196..215
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    718..839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   888..1017
                     /label=AAV2 ITR
     rep_origin      complement(1111..1566)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1248..1269)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1460..1479
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1593..1697
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1698..1766,1767..2558)
                     /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     complement(1916..1935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2729..3317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3218..3237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3586..3720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4145..4165)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4295..4360
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4780..5574)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4891..4910)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5501..5520
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5931..6260)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5945..6080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6245..6265
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gccggatcca aggtcgggca ggaagagggc ctatttccca tgattccttc atatttgcat
       61 atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata
      121 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      181 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      241 gctttatata tcttgtggaa aggacgaaac accgtgctcg cttcggcagc acatatacta
      301 gtcgacgggc cgcactcgcc ggtcccaagc ccggataaaa tgggaggggg cgggaaaccg
      361 cctaaccatg ccgagtgcgg ccgctgtggg aggaacctcc gcgaaagcgg tgaaggagag
      421 gcgcaaggtt aaccgcctca ggttccagga ggattcgtcc tccaaaggcc tccgcgaaag
      481 cggtgaagga gcgccacaag gttaactggc gcaggcctta cccacagtgg ccgcggtcgg
      541 cgtggactgt agaacactgc caatgccggt cccaagcccg gataaaagtg gagggtacag
      601 tccacgctct agagcggact tcggtccgct ttttactagg acctgcaggc atgcaagctt
      661 gacgtcggtt accgatatcc atatggcgac cgcatcgatc tcgagccgag gactagtaac
      721 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
      781 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
      841 catgtcttac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
      901 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
      961 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagaga
     1021 gggacagatc tgcagatctg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1081 caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat
     1141 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1201 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1261 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1321 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1381 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1441 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1501 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1561 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1621 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1681 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1741 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1801 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1861 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1921 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1981 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2041 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2101 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2161 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2221 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2281 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2341 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2401 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2461 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2521 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2581 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2641 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2701 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2761 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2821 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2881 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2941 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3001 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3061 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3121 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3181 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3241 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3301 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3361 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3421 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3481 tgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct ggaggggtgg
     3541 agtcgtgacg tgaattacgt catagggtta gggaggtcct ggatcgatcc agacatgata
     3601 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3661 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3721 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     3781 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatctcta gtcaaggcac
     3841 tatacatcaa atattcctta ttaacccctt tacaaattaa aaagctaaag gtacacaatt
     3901 tttgagcata gttattaata gcagacactc tatgcctgtg tggagtaaga aaaaacagta
     3961 tgttatgatt ataactgtta tgcctactta taaaggttac agaatatttt tccataattt
     4021 tcttgtatag cagtgcagct ttttcctttg tggtgtaaat agcaaagcaa gcaagagttc
     4081 tattactaaa cacagcatga ctcaaaaaac ttagcaattc tgaaggaaag tccttggggt
     4141 cttctacctt tctcttcttt tttggaggag tagaatgttg agagtcagca gtagcctcat
     4201 catcactaga tggcatttct tctgagcaaa acaggttttc ctcattaaag gcattccacc
     4261 actgctccca ttcatcagtt ccataggttg gaatctaaaa tacacaaaca attagaatca
     4321 gtagtttaac acattataca cttaaaaatt ttatatttac cttagagctt taaatctctg
     4381 taggtagttt gtccaattat gtcacaccac agaagtaagg ttccttcaca aagatccgga
     4441 ccaaagcggc catcgtgcct ccccactcct gcagttcggg ggcatggatg cgcggatagc
     4501 cgctgctggt ttcctggatg ccgacggatt tgcactgccg gtagaactcc gcgaggtcgt
     4561 ccagcctcag gcagcagctg aaccaactcg cgaggggatc gagcccgggg tgggcgaaga
     4621 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     4681 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg
     4741 gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag
     4801 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     4861 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     4921 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     4981 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     5041 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     5101 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     5161 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     5221 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     5281 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     5341 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
     5401 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     5461 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     5521 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
     5581 cgatcctcat cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg
     5641 caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc
     5701 tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag
     5761 cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca
     5821 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg
     5881 cttcctttag cagcccttgc gccctgagtg cttgcggcag cgtgaagctt tttgcaaaag
     5941 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     6001 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     6061 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     6121 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     6181 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     6241 ccacacccta actgacacac attccaca
//
