LOCUS       sequence-431907-        4244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AC17-5c riboswitch-regulated GFP reporter. The gene is inserted into
            downstream of T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4244)
  AUTHORS   Ishii Y, Fukunaga K, Cooney A, Yokobayashi Y, Matsuura T
  TITLE     Switchable and orthogonal gene expression control inside artificial 
            cells by synthetic riboswitches.
  JOURNAL   Chem Commun (Camb). 2024 Jun 4;60(46):5972-5975. doi: 
            10.1039/d4cc00965g.
  PUBMED    38767578
REFERENCE   2  (bases 1 to 4244)
  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"; journalName: 
            "Chem Commun (Camb)"; date: "2024-06-4- 4"; volume: "60"; issue: 
            "46"; pages: "5972-5975"
COMMENT     Sequence Label: pIVEX2.3-AC17-5c-frGFP
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(324..371)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     367..385
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             1222..1230
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1295..1314)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1296..1314)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1398..1417)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1537..1554)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1537..1553)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1546..1568)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1762..1781)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1881..1903
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1941..1959)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2027..2131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2132..2200,2201..2992)
                     /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(2350..2369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3163..3751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3652..3671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3905..3922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4039..4060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4075..4080,4081..4098,4099..4105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4113..4129
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4118..4140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4137..4153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4137..4153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        join(4174..4179,4180..4196,4197..4202)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg
       61 taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc gtccattccg
      121 acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg caatttctat
      181 gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt
      241 cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc ctgtggatat
      301 ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg
      361 ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg
      421 ttagcagccg gatccactag taacggccgc cagtgtgctg gaattcgccc ttttattatt
      481 tatacagctc gtccataccg tgcgtaatgc cagctgccgt aacaaactcc agtaagacca
      541 tgtggtcgcg tttttcgttc gggtctttgg acaaggctga ttgcgtagac aggtaatggt
      601 tgtccggaag caaaacaggt ccgtcaccaa tcggtgtatt ctgctggtaa tgatcggcaa
      661 gttggacaga accgtcttcg atgttatggc gaattttgaa gttcgctttg atgccgttct
      721 tctgcttatc cgcggtgata tacacgttgt ggctgttgta gttgtactcg agtttatggc
      781 ccagaatatt accgtcctcc ttaaagtcga tccctttgag ttcaatccga ttcaccagag
      841 tatcaccttc gaatttgact tctgcacggg ttttgtagtt accgtcgtct ttaaacgaaa
      901 tggtacgttc ctgcacgtaa ccttctggca tagcagattt aaagaagtcg tgacgtttca
      961 tgtgatcggg ataacggcta aagcactgta caccgtaagt cagggtagtc accagcgtag
     1021 gccaaggtac cggcagttta ccggttgtac agatgaattt cagggttaac ttaccgtaag
     1081 tagcgtctcc ttcaccttca ccggaaacac tgaatttgtg accgttaacg tcaccgtcta
     1141 actccaccag aatcggaact acaccagtaa acagttcttc gcctttcgac atttggatat
     1201 cgttcacgaa ttccattttt tacctcctta attcgataac cgggtcgcat caaagaggca
     1261 gcgtgaattc gcggacggaa tccgtctctc tcacccctat agtgagtcgt attaatttcg
     1321 cgggatcgag atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     1381 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     1441 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     1501 actgttgggc gccatctcct tgcatgcaag cttggcactg gccgtcgttt tacaacgtcg
     1561 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     1621 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     1681 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     1741 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     1801 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     1861 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     1921 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     1981 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2041 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2101 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2161 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2221 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2281 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2341 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2401 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2461 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2521 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2581 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2641 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2701 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2761 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2821 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2881 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2941 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3001 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3061 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3301 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3481 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3781 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3841 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3901 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3961 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     4021 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     4081 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     4141 aaacagctat gacatgatta cgaattctca tgtttgacag cttatcatcg ataagcttta
     4201 atgcggtagt ttatcacagt taaattgcta acgcagtcag gcac
//
