LOCUS       sequence-353898-        3126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  scRNA targeting J3 promoter for activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3126)
  AUTHORS   Tickman BI, Burbano DA, Chavali VP, Kiattisewee C, Fontana J, 
            Khakimzhan A, Noireaux V, Zalatan JG, Carothers JM
  TITLE     Multi-layer CRISPRa/i circuits for dynamic genetic programs in 
            cell-free and bacterial systems.
  JOURNAL   Cell Syst. 2021 Nov 17. pii: S2405-4712(21)00419-1. doi: 
            10.1016/j.cels.2021.10.008.
  PUBMED    34800362
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: "Cell Syst. 
            2021 Nov 17. pii: S2405-4712(21)00419-1. doi: 
            10.1016/j.cels.2021.10.008."
COMMENT     Sequence Label: pBT009.J1.J306
FEATURES             Location/Qualifiers
     source          1..3126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(19..607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(99..118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(778..1569,1570..1638))
                     /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"
     primer_bind     1401..1420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1639..1743)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1811..1829
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2351..2385
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     misc_RNA        2469..2487
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(2974..2991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
       61 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1381 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1741 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     1801 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct ggcgaatcct
     1861 ctgaccagcc agaaaacgac ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca
     1921 gcaagtgggg gacagcagaa gacctgaccg ccgcagagtg gatgtttgac atggtgaaga
     1981 ctatcgcacc atcagccaga aaaccgaatt ttgctgggtg ggctaacgat atccgcctga
     2041 tgcgtgaacg tgacggacgt aaccaccgcg acatgtgtgt gctgttccgc tgggcatgcc
     2101 aggacaactt ctggtccggt aacgtgctga gcccggccaa gcttagcccg aaaagtgcca
     2161 cctgtggcaa ttccgacgtc gcctacggta tccaccggag acctatggca gcctccggcc
     2221 gccataggac acctttggtt gccaagggtg acctatggtg accatgggcc accacgggcg
     2281 acctcaggta tcctgcggtg tcctgcggtt accaaaggcg tcctttgggt tccaccggat
     2341 acctccggac ttgacagcta gctcagtcct agggattgtg ctagcttgtg tccagaacgc
     2401 tccgtgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     2461 aaagtggcac atgaggatca cccatgtgct ttttttaacg catgagaaag cccccggaag
     2521 atcaccttcc gggggctttt ttattgcgcg gatccttact ccgtcgacaa gcttgcggcc
     2581 gcactcgagg aattccgact caattagttc agtcagtttc aggatattag tcatctctac
     2641 attgattatg agtattcaga aattccttaa atattctgac aaatgctctt tccctaaact
     2701 ccccccataa aaaaacccgc cgaagcgggt ttttacgtta tttgcggatt aacgattact
     2761 cgttatcaga accgcccaga cctgcgttca gcagttctgc caggctggca gatgcgtctt
     2821 ccggaattga tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     2881 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     2941 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3001 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3061 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3121 taaaaa
//
