LOCUS       Exported                2839 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pBAD-W108 scRNA.b1.
ACCESSION   .
VERSION     .
KEYWORDS    pCD315
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2839)
  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 2839)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2839
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     221..240
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        381..399
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     CDS             422..451
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             467..484
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(540..557)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      710..756
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(809..826)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(980..1568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1060..1079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1739..2599)
                     /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"
     primer_bind     2362..2381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2600..2704)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2772..2790
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2794..2837
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattctaaa gatctgaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
       61 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
      121 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
      181 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
      241 ataagattag cggattctac ctgacgcttt ttatcgcaac tctctactgt actagtgaag
      301 atccggcctg cagccagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
      361 atcaacttga aaaagtgcgc acatgaggat cacccatgtg ctttttttga agcttgggcc
      421 cgaacaaaaa ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca
      481 tcattgagtt taaacggtct ccagcttggc tgttttggcg gatgagagaa gattttcagc
      541 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
      601 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
      661 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
      721 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaactggat
      781 ccttactcga gtctagactg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
      841 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      901 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      961 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1021 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1081 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1141 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1201 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1261 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1321 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1381 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     1441 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1501 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1561 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1621 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1681 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1741 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1801 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1861 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1921 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1981 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2041 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2101 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2161 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2221 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2281 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2341 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2401 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2461 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2521 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2581 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2641 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     2701 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     2761 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     2821 tcgctaagga tgatttctg
//
