LOCUS       sequence-264425-        3024 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Malachite green aptamer with stability hairpin, bacterial promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3024)
  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 3024)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pJBL7007
FEATURES             Location/Qualifiers
     source          1..3024
                     /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"
     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(914..932)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      928..975
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1051..1067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1051..1067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1064..1086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1075..1091
                     /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(1099..1105,1106..1123,1124..1129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1144..1165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1282..1299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1453..2041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1533..1552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2212..3000)
                     /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     2835..2854
                     /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 actagtacgt cgactctcga gtgagattgt
      721 tgacggtacc gtattttggg atcccgactg gcgagagcca ggtaacgaat ggatcgggtc
      781 ggcatggcat ctccacctcc tcgcggtccg acctgggcat ccgaaggagg acgtcgtcca
      841 ctcggatggc taagggagag ctcggatccg gctgctaaca aagcccgaaa ggaagctgag
      901 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
      961 ttgaggggtt ttttgctgaa aggaggaact atatccggat cgagatcctc tagagtcgac
     1021 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1081 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1141 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1201 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1261 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1321 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1381 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1981 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2041 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2101 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2161 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2221 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2281 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2341 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2401 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2461 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2521 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2581 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2641 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2701 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2761 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2821 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2881 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2941 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3001 gtgagcaaaa acaggaaggc aaaa
//
