LOCUS       sequence-264426-        2983 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Malachite green aptamer, bacterial promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2983)
  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 2983)
  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: pJBL7006
FEATURES             Location/Qualifiers
     source          1..2983
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(49..153)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     221..239
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(277..299)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(394..1014)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDLDPDIVPPFSKKPLKTRLEAPRGYASYC
                     SAVAAANSASFRALLAAGSELSLSHPSGRRPPSDAQVGPRGGGDAMPTRSIRYLALASR
                     DPTSIIPRTELAVKVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTD
                     RPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     primer_bind     399..418
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     612..634
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     626..643
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     627..643
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        662..696
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(873..891)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      887..934
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1010..1026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1010..1026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1023..1045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1034..1050
                     /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(1058..1064,1065..1082,1083..1088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1103..1124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1241..1258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1412..2000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1492..1511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2171..2959)
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     2794..2813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
       61 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
      121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
      181 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      241 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      301 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      361 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      421 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      481 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
      541 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
      601 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg agctcggtac
      661 cttgacagct agctcagtcc taggtataat actagtggga tcccgactgg cgagagccag
      721 gtaacgaatg gatcgggtcg gcatggcatc tccacctcct cgcggtccga cctgggcatc
      781 cgaaggagga cgtcgtccac tcggatggct aagggagagc tcggatccgg ctgctaacaa
      841 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
      901 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatc
      961 gagatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt
     1021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2461 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2581 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaa
//
