LOCUS       sequence-409057-        3289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23106.EL222.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3289)
  AUTHORS   Alba Burbano D, Cardiff RAL, Tickman BI, Kiattisewee C, Maranas CJ, 
            Zalatan JG, Carothers JM
  TITLE     Engineering activatable promoters for scalable and multi-input 
            CRISPRa/i circuits.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Jul 25;120(30):e2220358120. doi: 
            10.1073/pnas.2220358120. Epub 2023 Jul 18.
  PUBMED    37463216
REFERENCE   2  (bases 1 to 3289)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2023-07-25"; volume: "120(30):e2220358120. 
            doi"; pages: " 10.1073/pnas.2220358120. Epub 2023 Jul 18"
COMMENT     Sequence Label: pDA010.EL222
FEATURES             Location/Qualifiers
     source          1..3289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     421..440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(659..763)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        790..824
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             831..842
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1559..1630
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1646..1673
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2157..2174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2328..2916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2408..2427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3087..3289,1..589,590..658))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
       61 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
      121 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      181 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      241 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      301 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      361 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
      421 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
      481 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
      541 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
      601 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
      661 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
      721 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      781 tgccacctgt ttacggctag ctcagtccta ggtatagtgc tagcctagag aaagaggaga
      841 aatactagat gttggatatg ggacaagatc ggccgatcga tggaagtggg gcacccgggg
      901 cagacgacac acgcgttgag gtgcaaccgc cggcgcagtg ggtcctcgac ctgatcgagg
      961 ccagcccgat cgcatcggtc gtgtccgatc cgcgtctcgc cgacaatccg ctgatcgcca
     1021 tcaaccaggc cttcaccgac ctgaccggct attccgaaga agaatgcgtc ggccgcaatt
     1081 gccgattcct ggcaggttcc ggcaccgagc cgtggctgac cgacaagatc cgccaaggcg
     1141 tgcgcgagca caagccggtg ctggtcgaga tcctgaacta caagaaggac ggcacgccgt
     1201 tccgcaatgc cgtgctcgtt gcaccgatct acgatgacga cgacgagctt ctctatttcc
     1261 tcggcagcca ggtcgaagtc gacgacgacc agcccaacat gggcatggcg cgccgcgaac
     1321 gcgccgcgga aatgctcaag acgctgtcgc cgcgccagct cgaggttacg acgctggtgg
     1381 catcgggctt gcgcaacaag gaagtggcgg cccggctcgg cctgtcggag aaaaccgtca
     1441 agatgcaccg cgggctggtg atggaaaagc tcaacctgaa gaccagcgcc gatctggtgc
     1501 gcattgccgt cgaagccgga atctaaggat ccaaactcga gtaaggatct ccaggcatca
     1561 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     1621 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttataa
     1681 acgcatgaga aagcccccgg aagatcacct tccgggggct tttttattgc gcggatcctt
     1741 actccgtcga caagcttgcg gccgcactcg aggaattccg actcaattag ttcagtcagt
     1801 ttcaggatat tagtcatctc tacattgatt atgagtattc agaaattcct taaatattct
     1861 gacaaatgct ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg
     1921 ttatttgcgg attaacgatt actcgttatc agaaccgccc agacctgcgt tcagcagttc
     1981 tgccaggctg gcagatgcgt cttccggaat tgatccgtcg accgatgccc ttgagagcct
     2041 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2101 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg
     2161 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2221 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2281 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2341 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2401 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2461 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2521 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2581 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2641 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2701 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2761 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2821 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2881 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2941 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3001 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3061 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3121 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3181 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3241 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagc
//
