LOCUS       sequence-226746-        3263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTet-W108 scRNA.b2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3263)
  AUTHORS   Dong C, Fontana J, Patel A, Carothers JM, Zalatan JG
  TITLE     Synthetic CRISPR-Cas gene activators for transcriptional 
            reprogramming in bacteria.
  JOURNAL   Nat Commun. 2018 Jun 27;9(1):2489. doi: 10.1038/s41467-018-04901-6.
  PUBMED    29950558
REFERENCE   2  (bases 1 to 3263)
  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: "Nat 
            Commun."; date: "2018-06-27"; volume: "9(1):2489. doi"; pages: " 
            10.1038/s41467-018-04901-6"
COMMENT     Sequence Label: pJF121
FEATURES             Location/Qualifiers
     source          1..3263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     582..601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(820..924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     992..1010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1014..1057)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(1075..1701)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     1606..1625
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1717..1772
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1753..1771
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        1864..1882
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     CDS             1905..1934
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1950..1967
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2023..2040)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2193..2239
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2292..2309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2463..3051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2543..2562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3222..3263,1..750,751..819))
                     /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"
ORIGIN
        1 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
       61 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
      121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
      181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
      241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
      421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
      481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
      541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
      601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
      661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
      721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
      781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
      961 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag
     1021 ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattctaaag atctttaaga
     1081 cccactttca catttaagtt gtttttctaa tccgcatatg atcaattcaa ggccgaataa
     1141 gaaggctggc tctgcacctt ggtgatcaaa taattcgata gcttgtcgta ataatggcgg
     1201 catactatca gtagtaggtg tttccctttc ttctttagcg acttgatgct cttgatcttc
     1261 caatacgcaa cctaaagtaa aatgccccac agcgctgagt gcatataatg cattctctag
     1321 tgaaaaacct tgttggcata aaaaggctaa ttgattttcg agagtttcat actgtttttc
     1381 tgtaggccgt gtacctaaat gtacttttgc tccatcgcga tgacttagta aagcacatct
     1441 aaaactttta gcgttattac gtaaaaaatc ttgccagctt tccccttcta aagggcaaaa
     1501 gtgagtatgg tgcctatcta acatctcaat ggctaaggcg tcgagcaaag cccgcttatt
     1561 ttttacatgc caatacaatg taggctgctc tacacctagc ttctgggcga gtttacgggt
     1621 tgttaaacct tcgattccga cctcattaag cagctctaat gcgctgttaa tcactttact
     1681 tttatctaat ctagacatca ttaattccta atttttgttg acactctatc gttgatagag
     1741 ttattttacc actccctatc agtgatagag aaaaactagt gaagatccgg cctgcagcca
     1801 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     1861 ggcacatgag gatcacccat gtgctttttt tgaagcttgg gcccgaacaa aaactcatct
     1921 cagaagagga tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttaaacgg
     1981 tctccagctt ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc
     2041 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
     2101 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
     2161 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     2221 actgggcctt tcgttttatc tgttgtttgt cggtgaactg gatccttact cgagtctaga
     2281 ctgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2341 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2401 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2461 tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2641 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2881 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3241 gaggcaccta tctcagcgat ctg
//
