LOCUS       sequence-272121-        2982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Malachite green aptamer, bacterial promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2982)
  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 2982)
  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..2982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(45..665)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDLDPDIVPPFSKKPLKTRLEAPRGYASYC
                     SAVAAANSASFRALLAAGSELSLSHPSGRRPPSDAQVGPRGGGDAMPTRSIRYLALASR
                     DPTSIIPRTELAVKVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTD
                     RPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     primer_bind     50..69
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     263..285
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     277..294
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     278..294
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        313..347
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(524..542)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      538..585
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(661..677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(661..677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(674..696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    685..701
                     /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(709..715,716..733,734..739))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    754..775
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(892..909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1062..1650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1142..1161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1821..2612,2613..2681))
                     /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     2444..2463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2682..2786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2854..2872
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2910..2932)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
       61 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
      121 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
      181 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
      241 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc
      301 gagctcggta ccttgacagc tagctcagtc ctaggtataa tactagtggg atcccgactg
      361 gcgagagcca ggtaacgaat ggatcgggtc ggcatggcat ctccacctcc tcgcggtccg
      421 acctgggcat ccgaaggagg acgtcgtcca ctcggatggc taagggagag ctcggatccg
      481 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
      541 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
      601 atatccggat cgagatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
      661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      721 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      781 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1021 gcaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1501 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1681 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1741 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1801 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2101 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2641 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2701 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2761 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2821 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     2881 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2941 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gt
//
