LOCUS       sequence-138508-        3425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AD1 Sense Expression Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3425)
  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 3425)
  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: pJBL2801
FEATURES             Location/Qualifiers
     source          1..3425
                     /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(164..166,167..880)
                     /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             164..877
                     /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(194..222)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     817..837
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             945..962
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1018..1035)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1188..1234
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1387..2046
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1453..1472)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2067..2161
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2568..2585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2702..3247)
                     /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(2782..2801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt agtttttaca
       61 gtgaattgtt ttaattagtt gtataaatgt tggagcagcg gggaatgtat acagttcatg
      121 tatatattcc ccgctttttt tttggatcta ggaggaagga tctatgagca aaggagaaga
      181 acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa
      241 attttctgtc cgtggagagg gtgaaggtga tgctacaaac ggaaaactca cccttaaatt
      301 tatttgcact actggaaaac tacctgttcc gtggccaaca cttgtcacta ctctgaccta
      361 tggtgttcaa tgcttttccc gttatccgga tcacatgaaa cggcatgact ttttcaagag
      421 tgccatgccc gaaggttatg tacaggaacg cactatatct ttcaaagatg acgggaccta
      481 caagacgcgt gctgaagtca agtttgaagg tgataccctt gttaatcgta tcgagttaaa
      541 gggtattgat tttaaagaag atggaaacat tcttggacac aaactcgagt acaactttaa
      601 ctcacacaat gtatacatca cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa
      661 aattcgccac aacgttgaag atggttccgt tcaactagca gaccattatc aacaaaatac
      721 tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcga cacaatctgt
      781 cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc
      841 tgctgggatt acacatggca tggatgagct ctacaaataa ggatctgaag cttgggccct
      901 aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc
      961 attgagttta aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct
     1021 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     1081 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     1141 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     1201 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aactggatcc
     1261 ttactcgagt ctagactgca gttgatcggg cacgtaagag gttccaactt tcaccataat
     1321 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     1381 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1441 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1501 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1561 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
     1621 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1681 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1741 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1801 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     1861 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1921 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1981 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     2041 gcgtaatttg atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga
     2101 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     2161 tccaagcctc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc
     2221 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc
     2281 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
     2341 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg
     2401 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta
     2461 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
     2521 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
     2581 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     2641 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     2701 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     2761 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     2821 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     2881 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     2941 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3001 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     3061 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     3121 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     3181 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     3241 atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat
     3301 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgttt
     3361 gacagcttat catcgataag cttccgatgg cgcgccgaga ggctttacac tttatgcttc
     3421 cggct
//
