LOCUS       sequence-254703-        6243 bp    DNA     circular SYN 14-APR-2025
DEFINITION  circular TAR Variant-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6243)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR
  TITLE     Live imaging of mRNA using RNA-stabilized fluorogenic proteins.
  JOURNAL   Nat Methods. 2019 Sep;16(9):862-865. doi: 10.1038/s41592-019-0531-7.
            Epub 2019 Aug 30.
  PUBMED    31471614
REFERENCE   2  (bases 1 to 6243)
  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/s41592-019-0531-7"; journalName: "Nat Methods"; date: 
            "2019-09"; volume: "16"; issue: "9"; pages: "862-865"
COMMENT     Sequence Label: pAV-U6+27-Tornado-F30-TAR Variant-1
FEATURES             Location/Qualifiers
     source          1..6243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      183..318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             689..1483
                     /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(743..762)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1353..1372
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     intron          1903..1968
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2098..2118
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2543..2677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2946..3534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3026..3045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3705..4496,4497..4565))
                     /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     4328..4347
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4566..4670)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4697..5152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4784..4803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4994..5015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     repeat_region   5246..5375
                     /label=AAV2 ITR
     polyA_signal    5424..5545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5965..6213)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6023..6042)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6193..6213)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(join(6241..6243,1..18))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
       61 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
      121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
      181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
      241 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
      301 gaggcttttt tggaggccta ggcttttgca aaaagcttca cgctgccgca agcactcagg
      361 gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg
      421 accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag
      481 aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac
      541 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
      601 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
      661 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
      721 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
      781 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
      841 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
      901 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
      961 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     1021 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     1081 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     1141 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     1201 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     1261 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     1321 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     1381 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     1441 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     1501 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     1561 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     1621 cgcggggatc tcatgctgga gttcttcgcc caccccgggc tcgatcccct cgcgagttgg
     1681 ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg caaatccgtc
     1741 ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact gcaggagtgg
     1801 ggaggcacga tggccgcttt ggtccggatc tttgtgaagg aaccttactt ctgtggtgtg
     1861 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
     1921 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
     1981 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
     2041 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
     2101 aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg
     2161 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
     2221 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
     2281 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
     2341 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
     2401 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
     2461 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2521 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2581 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2641 gtccaaactc atcaatgtat cttatcatgt ctggatcgat ccaggacctc cctaacccta
     2701 tgacgtaatt cacgtcacga ctccacccct ccaggaaccc ctagtgatgg agttggccac
     2761 tccctctctg cgcgctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2821 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2881 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2941 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3001 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3061 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3121 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3181 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3241 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3301 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3361 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3421 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3481 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3541 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3601 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3661 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3721 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3781 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3841 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3901 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3961 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4021 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4081 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4141 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4201 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4261 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4321 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4381 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4441 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4501 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4561 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4621 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4681 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4741 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4801 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4861 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4921 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4981 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5041 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5101 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc
     5161 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5221 agcagatctg cagatctgtc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc
     5281 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     5341 agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg
     5401 ctacttatct acgtaagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5461 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5521 cattataagc tgcaataaac aagttactag tcctcggctc gagatcgatg cggtcgccat
     5581 atggatatcg gtaaccgacg tcaagcttgc atgcctgcag gtcctagtaa aaagcggacc
     5641 gaagtccgct ctagagcgtg gactgtaccc tccactttta tccgggcttg ggaccggcat
     5701 tggcagtgtt ctacagtcca cgccgaccgc ggccgcttgc catgaatgat ccggtacctc
     5761 gggctcggag ctaatgagct cagacgagcc tttgctagcg gatcatcaga gtatgtggga
     5821 aacgtggcaa cgcgtcccac atacacatgg caagcggccg cactcggcat ggttaggcgg
     5881 tttcccgccc cctcccattt tatccgggct tgggaccggc gagtgcggcc cgtcgactag
     5941 tatatgtgct gccgaagcga gcacggtgtt tcgtcctttc cacaagatat ataaagccaa
     6001 gaaatcgaaa tactttcaag ttacggtaag catatgatag tccattttaa aacataattt
     6061 taaaactgca aactacccaa gaaattatta ctttctacgt cacgtatttt gtactaatat
     6121 ctttgtgttt acagtcaaat taattctaat tatctctcta acagccttgt atcgtatatg
     6181 caaatatgaa ggaatcatgg gaaataggcc ctcttcctgc ccgaccttgg atccggctgt
     6241 gga
//
