LOCUS       sequence-127162-        3500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pT181 Sense Expression Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3500)
  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 3500)
  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: pJBL2148
FEATURES             Location/Qualifiers
     source          1..3500
                     /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(239..241,242..955)
                     /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             239..952
                     /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(269..297)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     892..912
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             975..1004
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1020..1037
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1093..1110)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1263..1309
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1462..2121
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1528..1547)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2142..2236
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2643..2660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2777..3322)
                     /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(2857..2876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt acataaagat
       61 atatatttgg gtgagcgatt ccttaaacga aattgagatt aaggagtcgc tctttttttt
      121 ttatgtataa aaacaatcat gcaaatcatt caaatcattt ggaaaatcac gatttagaca
      181 atttttctaa aaccggctac tctaatagcc ggttgtaagg atctaggagg aaggatctat
      241 gagcaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
      301 tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa ggtgatgcta caaacggaaa
      361 actcaccctt aaatttattt gcactactgg aaaactacct gttccgtggc caacacttgt
      421 cactactctg acctatggtg ttcaatgctt ttcccgttat ccggatcaca tgaaacggca
      481 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa
      541 agatgacggg acctacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa
      601 tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattcttg gacacaaact
      661 cgagtacaac tttaactcac acaatgtata catcacggca gacaaacaaa agaatggaat
      721 caaagctaac ttcaaaattc gccacaacgt tgaagatggt tccgttcaac tagcagacca
      781 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
      841 gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct
      901 tgagtttgta actgctgctg ggattacaca tggcatggat gagctctaca aataaggatc
      961 tgaagcttgg gcccgaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc
     1021 atcatcatca tcatcattga gtttaaacgg tctccagctt ggctgttttg gcggatgaga
     1081 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     1141 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     1201 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     1261 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     1321 cggtgaactg gatccttact cgagtctaga ctgcagttga tcgggcacgt aagaggttcc
     1381 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     1441 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     1501 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     1561 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     1621 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt
     1681 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     1741 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     1801 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     1861 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     1921 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     1981 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     2041 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     2101 gagtggcagg gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc
     2161 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
     2221 ttttattggt gagaatccaa gcctccgatc aacgtctcat tttcgccaaa agttggccca
     2281 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     2341 cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac tgatttagtg
     2401 tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctcctgtt
     2461 cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctagcgga
     2521 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     2581 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     2641 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     2701 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     2761 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     2821 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2881 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     2941 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     3001 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     3061 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     3121 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     3181 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     3241 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     3301 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat
     3361 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     3421 gtaattctca tgtttgacag cttatcatcg ataagcttcc gatggcgcgc cgagaggctt
     3481 tacactttat gcttccggct
//
