LOCUS       sequence-409071-        3126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DA9 scRNA for activation of pDA303.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3126)
  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 3126)
  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: pBT009.119.DA9
FEATURES             Location/Qualifiers
     source          1..3126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     382..401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(620..724)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     792..810
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1332..1366
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        1450..1468
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(1955..1972)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2126..2714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2206..2225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2885..3126,1..550,551..619))
                     /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 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
       61 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      121 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      181 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      241 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      301 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      361 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
      421 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      481 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      541 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      601 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      661 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      721 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
      781 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc tggcgaatcc
      841 tctgaccagc cagaaaacga cctttctgtg gtgaaaccgg atgctgcaat tcagagcgcc
      901 agcaagtggg ggacagcaga agacctgacc gccgcagagt ggatgtttga catggtgaag
      961 actatcgcac catcagccag aaaaccgaat tttgctgggt gggctaacga tatccgcctg
     1021 atgcgtgaac gtgacggacg taaccaccgc gacatgtgtg tgctgttccg ctgggcatgc
     1081 caggacaact tctggtccgg taacgtgctg agcccggcca agcttagccc gaaaagttcc
     1141 acctgtggca attccgacgt cgcctacggt atccaccgga gacctatggc agcctccggc
     1201 cgtcatagga cacctttggt tgccaagggt gacctatggt gaccatgggc caccacgggc
     1261 gacctcaggt atcctgcggt gtcctgcggt taccaaaggc gtcctttggg ttccaccgga
     1321 tacctccgga cttgacagct agctcagtcc taggtataat actagttgca gccacgtgtg
     1381 agagttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     1441 aaaagtggca catgaggatc acccatgtgc tttttttaac gcatgagaaa gcccccggaa
     1501 gatcaccttc cgggggcttt tttattgcgc ggatccttac tccgtcgaca agcttgcggc
     1561 cgcactcgag gaattccgac tcaattagtt cagtcagttt caggatatta gtcatctcta
     1621 cattgattat gagtattcag aaattcctta aatattctga caaatgctct ttccctaaac
     1681 tccccccata aaaaaacccg ccgaagcggg tttttacgtt atttgcggat taacgattac
     1741 tcgttatcag aaccgcccag acctgcgttc agcagttctg ccaggctggc agatgcgtct
     1801 tccggaattg atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     1861 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     1921 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1981 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2041 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2101 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2161 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2221 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2281 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2341 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2401 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2461 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2521 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2581 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2641 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2701 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2761 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2821 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2881 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2941 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3001 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3061 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3121 tccagt
//
