LOCUS       sequence-489242-        2576 bp    DNA     circular SYN 14-OCT-2025
DEFINITION  Expression of an MS2 scRNA J306.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2576)
  AUTHORS   Kiattisewee C, Karanjia AV, Cardiff RAL, Olander KE, Leejareon P, 
            Alvi SS, Carothers JM, Zalatan JG
  TITLE     Systematic Mapping of Bacterial CRISPRa Systems for Synergistic Gene
            Activation Reveals Antagonistic Effects.
  JOURNAL   ACS Synth Biol. 2025 Jul 22. doi: 10.1021/acssynbio.5c00358.
  PUBMED    40693287
REFERENCE   2  (bases 1 to 2576)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2025 Jul 22. doi: 10.1021/acssynbio.5c00358."
COMMENT     Sequence Label: pCK303.306
FEATURES             Location/Qualifiers
     source          1..2576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(90..184)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(215..1009)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(326..345)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     936..955
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1188..1206
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1210..1253)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1271..1305
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        1389..1407
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     CDS             1430..1459
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1475..1492
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1548..1565)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1718..1764
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1817..1834)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1988..2576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2068..2087)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagagt actgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
      121 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
      181 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
      241 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
      301 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
      361 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
      421 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
      481 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
      541 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
      601 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
      661 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
      721 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
      781 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
      841 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
      901 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
      961 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     1021 ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga
     1081 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     1141 attccgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1201 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt
     1261 ctaaagatct ttgacagcta gctcagtcct aggtataata ctagtttgtg tccagaacgc
     1321 tccgtgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     1381 aaagtggcac atgaggatca cccatgtgct ttttttgaag cttgggcccg aacaaaaact
     1441 catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta
     1501 aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
     1561 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
     1621 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     1681 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     1741 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aactggatcc ttactcgagt
     1801 ctagactgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     1861 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1921 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1981 tggcgttttt ccacaggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2041 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2101 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2161 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2221 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2281 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2341 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2401 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2461 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2521 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaa
//
