LOCUS       sequence-127164-        3652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Anti-anti Sense Expression Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3652)
  AUTHORS   Chappell J, Takahashi MK, Lucks JB
  TITLE     Creating small transcription activating RNAs.
  JOURNAL   Nat Chem Biol. 2015 Mar;11(3):214-20. doi: 10.1038/nchembio.1737. 
            Epub 2015 Feb 2.
  PUBMED    25643173
REFERENCE   2  (bases 1 to 3652)
  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/nchembio.1737"; journalName: "Nat Chem Biol"; date: 
            "2015-03"; volume: "11"; issue: "3"; pages: "214-20"
COMMENT     Sequence Label: pJBL1864
FEATURES             Location/Qualifiers
     source          1..3652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..50
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(391..393,394..1107)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             391..1104
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(421..449)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1044..1064
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1127..1156
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1172..1189
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1245..1262)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1415..1461
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1614..2273
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1680..1699)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2294..2388
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2795..2812)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2929..3474)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3009..3028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt aacaaaatcg
       61 actccttttt cctatgtcta gtccacatca gaaaaaggag tcgctcacgc cctgaccaaa
      121 gtttgtgaac gacatcattc aaagaaaaaa acactgagtt gtttttataa tcttgtatat
      181 ttagatatta aacgatattt aaatatacat aaagatatat atttgggtga gcgattcctt
      241 aaacgaaatt gagattaagg agtcgctctt ttttatgtat aaaaacaatc atgcaaatca
      301 ttcaaatcat ttggaaaatc acgatttaga caatttttct aaaaccggct actctaatag
      361 ccggttgtaa ggatctagga ggaaggatct atgagcaaag gagaagaact tttcactgga
      421 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt
      481 ggagagggtg aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact
      541 ggaaaactac ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc
      601 ttttcccgtt atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa
      661 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct
      721 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt
      781 aaagaagatg gaaacattct tggacacaaa ctcgagtaca actttaactc acacaatgta
      841 tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac
      901 gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
      961 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat
     1021 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
     1081 catggcatgg atgagctcta caaataagga tctgaagctt gggcccgaac aaaaactcat
     1141 ctcagaagag gatctgaata gcgccgtcga ccatcatcat catcatcatt gagtttaaac
     1201 ggtctccagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
     1261 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
     1321 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     1381 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
     1441 agactgggcc tttcgtttta tctgttgttt gtcggtgaac tggatcctta ctcgagtcta
     1501 gactgcagtt gatcgggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     1561 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
     1621 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     1681 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     1741 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     1801 ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac ggtgagctgg
     1861 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     1921 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     1981 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     2041 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     2101 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     2161 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     2221 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taatttgata
     2281 tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     2341 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcctccga
     2401 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     2461 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg
     2521 tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt
     2581 ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg
     2641 caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta tgttggcact
     2701 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     2761 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     2821 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     2881 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     2941 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     3001 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     3061 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     3121 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     3181 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     3241 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     3301 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     3361 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     3421 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     3481 catcttatta atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta
     3541 gcacctgaag tcagccccat acgatataag ttgtaattct catgtttgac agcttatcat
     3601 cgataagctt ccgatggcgc gccgagaggc tttacacttt atgcttccgg ct
//
