LOCUS       sequence-272112-        3007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Malachite green aptamer with stability hairpin, T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3007)
  AUTHORS   Silverman AD, Kelley-Loughnane N, Lucks JB, Jewett MC
  TITLE     Deconstructing Cell-Free Extract Preparation for in Vitro Activation
            of Transcriptional Genetic Circuitry.
  JOURNAL   ACS Synth Biol. 2019 Feb 15;8(2):403-414. doi: 
            10.1021/acssynbio.8b00430. Epub 2019 Jan 29.
  PUBMED    30596483
REFERENCE   2  (bases 1 to 3007)
  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: 
            "10.1021/acssynbio.8b00430"; journalName: "ACS Synth Biol"; date: 
            "2019-02-15- 15"; volume: "8"; issue: "2"; pages: "403-414"
COMMENT     Sequence Label: pJBL7005
FEATURES             Location/Qualifiers
     source          1..3007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        36..54
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(272..290)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      286..333
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(409..425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(409..425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(422..444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    433..449
                     /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)."
     promoter        complement(join(457..463,464..481,482..487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    502..523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(640..657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(810..1398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(890..909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1569..2360,2361..2429))
                     /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     2192..2211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2430..2534)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2602..2620
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2658..2680)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2780..2799
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(2993..3007,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3007,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacctaata cgactcacta taggacgtcg
       61 actctcgagt gagattgttg acggtaccgt attttgggat cccgactggc gagagccagg
      121 taacgaatgg atcgggtcgg catggcatct ccacctcctc gcggtccgac ctgggcatcc
      181 gaaggaggac gtcgtccact cggatggcta agggagagct cggatccggc tgctaacaaa
      241 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
      301 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatcg
      361 agatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
      421 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      481 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
      541 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
      601 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      661 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      721 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaggccagg
      781 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      841 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      901 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      961 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1021 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1081 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1141 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1201 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1261 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1321 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1381 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1441 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1501 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1561 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1621 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1681 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1741 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1801 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1861 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     1921 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     1981 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2041 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2101 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2161 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2221 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2281 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2341 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2401 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2461 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2521 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     2581 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     2641 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     2701 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     2761 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     2821 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     2881 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     2941 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     3001 cgacgtt
//
