LOCUS       sequence-344821-        6264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of J1-BBa_J23117-mRFP and J109 scRNA on pBBR1-GmR 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6264)
  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 6264)
  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: pPPC016.109
FEATURES             Location/Qualifiers
     source          1..6264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(362..367,368..384,385..390))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     terminator      complement(556..583)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(599..670)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(697..1374)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             1381..1392
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(1402..1436)
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     protein_bind    1729..1750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1765..1770,1771..1788,1789..1795)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1803..1819
                     /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)."
     primer_bind     1808..1830
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1827..1843
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1827..1843
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1862..1882
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1864..1882
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(1927..1973)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2126..2143
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2199..2216)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2232..2261)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_RNA        complement(2284..2302)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        complement(2386..2420)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(2434..2453)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2435..2453)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2463..2480)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2463..2479)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2472..2494)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3188..3850)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      3851..4622
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             complement(join(5904..6264,1..173))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat
       61 ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac
      121 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct
      181 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
      241 catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc
      301 accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta
      361 cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg
      421 tgtgcgtcca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
      481 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
      541 cagtttttat ataaacgcag aaaggcccac ccgaaggtga gccagtgtga ctctagtaga
      601 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
      661 gttttatttg atgcctggag atccttactc gagtttttaa gcaccggtgg agtgacgacc
      721 ttcagcacgt tcgtactgtt caacgatggt gtagtcttcg ttgtgggagg tgatgtccag
      781 tttgatgtcg gttttgtaag cacccggcag ctgaaccggt tttttagcca tgtaggtggt
      841 tttaacttca gcgtcgtagt gaccaccgtc tttcagtttc agacgcattt tgatttcacc
      901 tttcagagca ccgtcttccg ggtacatacg ttcggtggaa gcttcccaac ccatggtttt
      961 tttctgcata accggaccgt cggacgggaa gttggtacca cgcagtttaa ctttgtagat
     1021 gaactcaccg tcttgcaggg aggagtcctg ggtaacggta acaacaccac cgtcttcgaa
     1081 gttcataaca cgttcccatt tgaaaccttc cgggaaggac agtttcaggt agtccgggat
     1141 gtcagccggg tgtttaacgt aagctttgga accgtactgg aactgcgggg acaggatgtc
     1201 ccaagcgaac ggcagcggac cacctttggt aactttcagt ttagcggtct gggtaccttc
     1261 gtacggacga ccttcacctt caccttcgat ttcgaactcg tgaccgttaa cggaaccttc
     1321 catacgaact ttgaaacgca tgaactcttt gataacgtct tcgctactcg ccatggtacc
     1381 tttctcctct ttaatgaatt cgctagcaca atccctagga ctgagctagc tgtcaagtcc
     1441 ggaggtatcc ggtggaaccc aaaggacgcc tttggtaacc gcaggacacc gcaggatacc
     1501 tgaggtcgcc cgtggtggcc catggtcacc ataggtcacc cttggcaacc aaaggtgtcc
     1561 tatggcggcc ggaggctgcc ataggtctcc ggtggatacc gtaggcgacg tcggaattgc
     1621 catgcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     1681 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     1741 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     1801 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
     1861 cgcaattaac cctcactaaa gggaacaaaa gctggtctag cttaagagtt caccgacaaa
     1921 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg
     1981 gcagttccct actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac
     2041 ttctgagttc ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg
     2101 ttttatcaga ccgcttctgc gttctgattt aatctgtatc aggctgaaaa tcttctctca
     2161 tccgccaaaa cagccaagct ggagaccgtt taaactcaat gatgatgatg atgatggtcg
     2221 acggcgctat tcagatcctc ttctgagatg agtttttgtt cgggcccaag cttcaaaaaa
     2281 agcacatggg tgatcctcat gtgccacttt ttcaagttga taacggacta gccttatttt
     2341 aacttgctat ttctagctct aaaaccagga caccgcagga tacctactag tattatacct
     2401 aggactgagc tagctgtcaa gagctccaat tcgccctata gtgagtcgta ttacgcgcgc
     2461 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2521 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2581 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt
     2641 tgttaaaatc cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgtac
     2701 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
     2761 cctggttgct acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgaaagca
     2821 aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat gtctattgct
     2881 ggtttaccgg tttattgact accggaagca gtgtgaccgt gtgcttctca aatgcctgag
     2941 gccagtttgc tcaggctctc cccgtggagg taataattga cgatatgatc atttattctg
     3001 cctcccagag cctgataaaa acggtgaatc cgttagcgag gtgccgccgg cttccattca
     3061 ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag
     3121 ggcggcgagg cggctacagc cgatagtctg gaacagcgca cttacgggtt gctgcgcaac
     3181 ccaagtgcta ccggcgcggc agcgtgaccc gtgtcggcgg ctccaacggc tcgccatcgt
     3241 ccagaaaaca cggctcatcg ggcatcggca ggcgctgctg cccgcgccgt tcccattcct
     3301 ccgtttcggt caaggctggc aggtctggtt ccatgcccgg aatgccgggc tggctgggcg
     3361 gctcctcgcc ggggccggtc ggtagttgct gctcgcccgg atacagggtc gggatgcggc
     3421 gcaggtcgcc atgccccaac agcgattcgt cctggtcgtc gtgatcaacc accacggcgg
     3481 cactgaacac cgacaggcgc aactggtcgc ggggctggcc ccacgccacg cggtcattga
     3541 ccacgtaggc cgacacggtg ccggggccgt tgagcttcac gacggagatc cagcgctcgg
     3601 ccaccaagtc cttgactgcg tattggaccg tccgcaaaga acgtccgatg agcttggaaa
     3661 gtgtcttctg gctgaccacc acggcgttct ggtggcccat ctgcgccacg aggtgatgca
     3721 gcagcattgc cgccgtgggt ttcctcgcaa taagcccggc ccacgcctca tgcgctttgc
     3781 gttccgtttg cacccagtga ccgggcttgt tcttggcttg aatgccgatt tctctggact
     3841 gcgtggccat gcttatctcc atgcggtagg ggtgccgcac ggttgcggca ccatgcgcaa
     3901 tcagctgcaa cttttcggca gcgcgacaac aattatgcgt tgcgtaaaag tggcagtcaa
     3961 ttacagattt tctttaacct acgcaatgag ctattgcggg gggtgccgca atgagctgtt
     4021 gcgtaccccc cttttttaag ttgttgattt ttaagtcttt cgcatttcgc cctatatcta
     4081 gttctttggt gcccaaagaa gggcacccct gcggggttcc cccacgcctt cggcgcggct
     4141 ccccctccgg caaaaagtgg cccctccggg gcttgttgat cgactgcgcg gccttcggcc
     4201 ttgcccaagg tggcgctgcc cccttggaac ccccgcactc gccgccgtga ggctcggggg
     4261 gcaggcgggc gggcttcgcc cttcgactgc ccccactcgc ataggcttgg gtcgttccag
     4321 gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc cttccacttg
     4381 gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc ccagagaaaa
     4441 ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg ggtatgtggt
     4501 cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag gcgcagcctg
     4561 tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca gccggtggcc
     4621 gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga ccgcgcaggg
     4681 cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc acgactttgc
     4741 gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc accgccttgc
     4801 gctgcccccg tcgagccggt tggacaccaa aagggagggg caggcatggc ggcatacgcg
     4861 atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg cggccagtct caagcacgcc
     4921 taccgcgagc gcgagacgcc caacgctgac gccagcagga cgccagagaa cgagcactgg
     4981 gcggccagca gcaccgatga agcgatgggc cgactgcgcg agttgctgcc agagaagcgg
     5041 cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg ccagcccgga atggtggaag
     5101 tcggccagcc aagaacagca ggcggcgttc ttcgagaagg cgcacaagtg gctggcggac
     5161 aagtacgggg cggatcgcat cgtgacggcc agcatccacc gtgacgaaac cagcccgcac
     5221 atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc tgtcggccaa ggagttcatc
     5281 ggcaacaaag cgcagatgac ccgcgaccag accacgtttg cggccgctgt ggccgatcta
     5341 gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca ggcgttctac
     5401 gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc ggtcgagcca
     5461 cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg cgttgagacg
     5521 ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga gcctgcccta
     5581 caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca agagacggcc
     5641 cgagaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt gaatcgggat
     5701 atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct gacggaacag
     5761 cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg cgcacatttc
     5821 cccgaaaagt gccacctgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
     5881 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
     5941 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
     6001 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
     6061 agcgcggtgg caatgccctg cctccggtgc tctccggaga ctgcgagatc atagatatag
     6121 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc
     6181 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
     6241 taatcggagt ccggctgatg ttgg
//
