LOCUS       sequence-272120-        2966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Malachite green aptamer, T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2966)
  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 2966)
  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: pJBL7004
FEATURES             Location/Qualifiers
     source          1..2966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..30)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     130..149
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(229..729)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDLDPDIVPPFSKKPLKTRLEAPRGYASYC
                     SAVAAANSASFRALLAAGSELSLSHPSGRRPPSDAQVGPRGGGDAMPTRSIRYLALASR
                     DPPIVSRIRYRARIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNS
                     CAA"
     primer_bind     343..365
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     357..374
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     358..374
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     393..412
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        393..411
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(588..606)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      602..649
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(725..741)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(725..741)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(738..760)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    749..765
                     /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(773..779,780..797,798..803))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    818..839
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(956..973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1126..1714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1206..1225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1885..2676,2677..2745))
                     /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     2508..2527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2746..2850)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2918..2936
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
       61 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
      121 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
      181 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
      241 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      301 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      361 aaacgacggc cagtgaattc gagctcggta cctaatacga ctcactatag ggggatcccg
      421 actggcgaga gccaggtaac gaatggatcg ggtcggcatg gcatctccac ctcctcgcgg
      481 tccgacctgg gcatccgaag gaggacgtcg tccactcgga tggctaaggg agagctcgga
      541 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
      601 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
      661 aactatatcc ggatcgagat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat
      721 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
      781 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
      841 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
      901 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
      961 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1021 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1081 gccagcaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2761 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2821 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2881 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     2941 tcgtctcgcg cgtttcggtg atgacg
//
