LOCUS       sequence-344728-        5195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of J106 scRNA on pBBR1-GmR plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5195)
  AUTHORS   Kiattisewee C, Dong C, Fontana J, Sugianto W, Peralta-Yahya P, 
            Carothers JM, Zalatan JG
  TITLE     Portable bacterial CRISPR transcriptional activation enables 
            metabolic engineering in Pseudomonas putida.
  JOURNAL   Metab Eng. 2021 Apr 27. pii: S1096-7176(21)00063-X. doi: 
            10.1016/j.ymben.2021.04.002.
  PUBMED    33930546
REFERENCE   2  (bases 1 to 5195)
  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: "Metab Eng. 
            2021 Apr 27. pii: S1096-7176(21)00063-X. doi: 
            10.1016/j.ymben.2021.04.002."
COMMENT     Sequence Label: pPPC008.106
FEATURES             Location/Qualifiers
     source          1..5195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             103..636
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1918..2689
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2690..3352
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     4046..4068
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4060..4077
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4061..4077
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4087..4106
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4087..4105
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4120..4154
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        4238..4256
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     CDS             4279..4308
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4324..4341
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4397..4414)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4567..4613
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4658..4678)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4658..4676)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4697..4713)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4697..4713)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4710..4732)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4721..4737
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4745..4751,4752..4769,4770..4775))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4790..4811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5081..5086,5087..5103,5104..5109)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tggcttgtta tgactgtttt tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc
       61 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagcaac
      121 gatgttacgc agcagggcag tcgccctaaa acaaagttag gtggctcaag tatgggcatc
      181 attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt
      241 ttcggtcgtg agttcggaga cgtagccacc tactcccaac atcagccgga ctccgattac
      301 ctcgggaact tgctccgtag taagacattc atcgcgcttg ctgccttcga ccaagaagcg
      361 gttgttggcg ctctcgcggc ttacgttctg cccaagtttg agcagccgcg tagtgagatc
      421 tatatctatg atctcgcagt ctccggagag caccggaggc agggcattgc caccgcgctc
      481 atcaatctcc tcaagcatga ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca
      541 gattacggtg acgatcccgc agtggctctc tatacaaagt tgggcatacg ggaagaagtg
      601 atgcactttg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc cgagatcggc
      661 ttcccggccg cggagttgtt cggtaaattg tcacaacgcc gcaggtggca cttttcgggg
      721 aaatgtgcgc gcccgcgttc ctgctggcgc tgggcctgtt tctggcgctg gacttcccgc
      781 tgttccgtca gcagcttttc gcccacggcc ttgatgatcg cggcggcctt ggcctgcata
      841 tcccgattca acggccccag ggcgtccaga acgggcttca ggcgctcccg aaggtctcgg
      901 gccgtctctt gggcttgatc ggccttcttg cgcatctcac gcgctcctgc ggcggcctgt
      961 agggcaggct catacccctg ccgaaccgct tttgtcagcc ggtcggccac ggcttccggc
     1021 gtctcaacgc gctttgagat tcccagcttt tcggccaatc cctgcggtgc ataggcgcgt
     1081 ggctcgaccg cttgcgggct gatggtgacg tggcccactg gtggccgctc cagggcctcg
     1141 tagaacgcct gaatgcgcgt gtgacgtgcc ttgctgccct cgatgccccg ttgcagccct
     1201 agatcggcca cagcggccgc aaacgtggtc tggtcgcggg tcatctgcgc tttgttgccg
     1261 atgaactcct tggccgacag cctgccgtcc tgcgtcagcg gcaccacgaa cgcggtcatg
     1321 tgcgggctgg tttcgtcacg gtggatgctg gccgtcacga tgcgatccgc cccgtacttg
     1381 tccgccagcc acttgtgcgc cttctcgaag aacgccgcct gctgttcttg gctggccgac
     1441 ttccaccatt ccgggctggc cgtcatgacg tactcgaccg ccaacacagc gtccttgcgc
     1501 cgcttctctg gcagcaactc gcgcagtcgg cccatcgctt catcggtgct gctggccgcc
     1561 cagtgctcgt tctctggcgt cctgctggcg tcagcgttgg gcgtctcgcg ctcgcggtag
     1621 gcgtgcttga gactggccgc cacgttgccc attttcgcca gcttcttgca tcgcatgatc
     1681 gcgtatgccg ccatgcctgc ccctcccttt tggtgtccaa ccggctcgac gggggcagcg
     1741 caaggcggtg cctccggcgg gccactcaat gcttgagtat actcactaga ctttgcttcg
     1801 caaagtcgtg accgcctacg gcggctgcgg cgccctacgg gcttgctctc cgggcttcgc
     1861 cctgcgcggt cgctgcgctc ccttgccagc ccgtggatat gtggacgatg gccgcgagcg
     1921 gccaccggct ggctcgcttc gctcggcccg tggacaaccc tgctggacaa gctgatggac
     1981 aggctgcgcc tgcccacgag cttgaccaca gggattgccc accggctacc cagccttcga
     2041 ccacataccc accggctcca actgcgcggc ctgcggcctt gccccatcaa tttttttaat
     2101 tttctctggg gaaaagcctc cggcctgcgg cctgcgcgct tcgcttgccg gttggacacc
     2161 aagtggaagg cgggtcaagg ctcgcgcagc gaccgcgcag cggcttggcc ttgacgcgcc
     2221 tggaacgacc caagcctatg cgagtggggg cagtcgaagg gcgaagcccg cccgcctgcc
     2281 ccccgagcct cacggcggcg agtgcggggg ttccaagggg gcagcgccac cttgggcaag
     2341 gccgaaggcc gcgcagtcga tcaacaagcc ccggaggggc cactttttgc cggaggggga
     2401 gccgcgccga aggcgtgggg gaaccccgca ggggtgccct tctttgggca ccaaagaact
     2461 agatataggg cgaaatgcga aagacttaaa aatcaacaac ttaaaaaagg ggggtacgca
     2521 acagctcatt gcggcacccc ccgcaatagc tcattgcgta ggttaaagaa aatctgtaat
     2581 tgactgccac ttttacgcaa cgcataattg ttgtcgcgct gccgaaaagt tgcagctgat
     2641 tgcgcatggt gccgcaaccg tgcggcaccc ctaccgcatg gagataagca tggccacgca
     2701 gtccagagaa atcggcattc aagccaagaa caagcccggt cactgggtgc aaacggaacg
     2761 caaagcgcat gaggcgtggg ccgggcttat tgcgaggaaa cccacggcgg caatgctgct
     2821 gcatcacctc gtggcgcaga tgggccacca gaacgccgtg gtggtcagcc agaagacact
     2881 ttccaagctc atcggacgtt ctttgcggac ggtccaatac gcagtcaagg acttggtggc
     2941 cgagcgctgg atctccgtcg tgaagctcaa cggccccggc accgtgtcgg cctacgtggt
     3001 caatgaccgc gtggcgtggg gccagccccg cgaccagttg cgcctgtcgg tgttcagtgc
     3061 cgccgtggtg gttgatcacg acgaccagga cgaatcgctg ttggggcatg gcgacctgcg
     3121 ccgcatcccg accctgtatc cgggcgagca gcaactaccg accggccccg gcgaggagcc
     3181 gcccagccag cccggcattc cgggcatgga accagacctg ccagccttga ccgaaacgga
     3241 ggaatgggaa cggcgcgggc agcagcgcct gccgatgccc gatgagccgt gttttctgga
     3301 cgatggcgag ccgttggagc cgccgacacg ggtcacgctg ccgcgccggt agcacttggg
     3361 ttgcgcagca acccgtaagt gcgctgttcc agactatcgg ctgtagccgc ctcgccgccc
     3421 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
     3481 gaatggaagc cggcggcacc tcgctaacgg attcaccgtt tttatcaggc tctgggaggc
     3541 agaataaatg atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc
     3601 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     3661 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     3721 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc
     3781 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     3841 tacggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3901 aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     3961 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     4021 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     4081 cgcgcgtaat acgactcact atagggcgaa ttggagctct tgacagctag ctcagtccta
     4141 ggtataatac tagtaggacg cctttggtaa ccgcgtttta gagctagaaa tagcaagtta
     4201 aaataaggct agtccgttat caacttgaaa aagtggcaca tgaggatcac ccatgtgctt
     4261 tttttgaagc ttgggcccga acaaaaactc atctcagaag aggatctgaa tagcgccgtc
     4321 gaccatcatc atcatcatca ttgagtttaa acggtctcca gcttggctgt tttggcggat
     4381 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     4441 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     4501 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     4561 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4621 ttgtcggtga actcttaagc tagaccagct tttgttccct ttagtgaggg ttaattgcgc
     4681 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4741 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4801 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4861 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgcatgc
     4921 ataaaaactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     4981 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     5041 tggacgcaca ccgtggaaac ggatgaaggc acgaacccag ttgacataag cctgttcggt
     5101 tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     5161 acgcagcggt ggtaacggcg cagtggcggt tttca
//
