LOCUS       sequence-461218-        5202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Cre recombinase activity and the PCB 
            production gene. Cre removes a transcription terminator, leading to 
            mCherry production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5202)
  AUTHORS   Jafarbeglou F, Dunlop MJ
  TITLE     Red Light Responsive Cre Recombinase for Bacterial Optogenetics.
  JOURNAL   ACS Synth Biol. 2024 Nov 19. doi: 10.1021/acssynbio.4c00388.
  PUBMED    39558834
REFERENCE   2  (bases 1 to 5202)
  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: "ACS Synth 
            Biol. 2024 Nov 19. doi: 10.1021/acssynbio.4c00388."
COMMENT     Sequence Label: pBbAk-w4-loxP-TT-loxP-mCherry-PCB
FEATURES             Location/Qualifiers
     source          1..5202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(164..197)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      245..316
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      332..359
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(368..401)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      1198..1269
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1285..1312
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1340..1357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1474..2019)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1554..1573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2069..2103
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             2120..2131
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             2863..2874
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3642..3713
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3729..3756
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3812..3906)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3937..4731)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4048..4067)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4658..4677
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4916..4993)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5199..5202,1..16)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
       61 ccgaagttcc tattatcacg aggccctttg agttatcaaa aagagtattg cattaaagtc
      121 taacctatag gaatcttaca gccatcgaga gggacacggc gaaataactt cgtatagcat
      181 acattatacg aagttatcca ggcatcaaat aaggatccaa actcgagtaa ggatctccag
      241 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      301 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
      361 ttataccata acttcgtata gcatacatta tacgaagtta ttttaagaag gagatataca
      421 tatgactagc aaaagatcca agggcgagga ggataacatg gctatcatta aagagttcat
      481 gcgcttcaaa gttcacatgg agggttctgt taacggtcac gagttcgaga tcgaaggcga
      541 aggcgagggc cgtccgtatg aaggcaccca gaccgccaaa ctgaaagtga ctaaaggcgg
      601 cccgctgcct tttgcgtggg acatcctgag cccgcaattt atgtacggtt ctaaagcgta
      661 tgttaaacac ccagcggata tcccggacta tctgaagctg tcttttccgg aaggtttcaa
      721 gtgggaacgc gtaatgaatt ttgaagatgg tggtgtcgtg accgtcactc aggactcctc
      781 cctgcaagat ggcgagttca tctataaagt taaactgcgt ggtactaatt ttccatctga
      841 tggcccggtg atgcagaaaa agacgatggg ttgggaggcg tctagcgaac gcatgtatcc
      901 ggaagatggt gcgctgaaag gcgaaattaa acagcgcctg aaactgaaag atggcggcca
      961 ttatgacgct gaagtgaaaa ccacgtacaa agccaagaaa cctgtgcagc tgcctggcgc
     1021 gtacaatgtg aatattaaac tggacatcac ctctcataat gaagattata cgatcgtaga
     1081 gcaatatgag cgcgcggagg gtcgtcattc taccggtggc atggatgagc tgtacaaata
     1141 atagggatct ccaggcatca aataaggatc caaactcgag taaggatctc caggcatcaa
     1201 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1261 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatacc
     1321 tagggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     1381 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     1441 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     1501 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     1561 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     1621 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     1681 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     1741 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     1801 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     1861 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     1921 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     1981 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     2041 ataaaatatt tctagatttc agtgcaatct gacagctagc tcagtcctag gtataatgct
     2101 agctcacaca gaattcatta aagaggagaa aggtaccatg agtgtcaact tagcttccca
     2161 gttgcgggaa gggacgaaaa aatcccactc catggcggag aacgtcggct ttgtcaaatg
     2221 cttcctcaag ggcgttgtcg agaaaaattc ctaccgtaag ctggttggca atctctactt
     2281 tgtctacagt gccatggaag aggaaatggc aaaatttaag gaccatccca tcctcagcca
     2341 catttacttc cccgaactca accgcaaaca aagcctagag caagacctgc aattctatta
     2401 cggctccaac tggcggcaag aagtgaaaat ttctgccgct ggccaagcct atgtggaccg
     2461 agtccggcaa gtggccgcta cggcccctga attgttggtg gcccattcct acacccgtta
     2521 cctgggggat ctttccggcg gtcaaattct caagaaaatt gcccaaaatg ccatgaatct
     2581 ccacgatggt ggcacagctt tctatgaatt tgccgacatt gatgacgaaa aggcttttaa
     2641 aaatacctac cgtcaagcta tgaatgatct gcccattgac caagccaccg ccgaacggat
     2701 tgtggatgaa gccaatgacg cctttgccat gaacatgaaa atgttcaacg aacttgaagg
     2761 caacctgatc aaggcgatcg gcattatggt gttcaacagc ctcacccgtc gccgcagtca
     2821 aggcagcacc gaagttggcc tcgccacctc cgaaggctag ttaaagagga gaaaggatcc
     2881 atggccgtca ctgatttaag tttgaccaat tcttccctga tgcccacgtt gaacccgatg
     2941 attcaacagt tggccctggc gatcgccgct agttggcaaa gtttacccct caagccctat
     3001 caattgccgg aggatttggg ctacgtagaa ggccgcctgg aaggggaaaa gttagtgatt
     3061 gaaaatcggt gctaccaaac gccccagttt cgcaaaatgc atttggagtt ggccaaggtg
     3121 ggcaaagggt tggatattct ccactgtgta atgtttcctg agcctttata cggtctacct
     3181 ttgtttggct gtgacattgt ggccggcccc ggtggagtaa gtgcggctat tgcggatcta
     3241 tcccccaccc aaagcgatcg ccaattgccc gcagcgtacc aaaaatcatt ggcagagcta
     3301 ggccagccag aatttgagca acaacgggaa ttgcccccct ggggagaaat attttctgaa
     3361 tattgtttat tcatccgtcc cagcaatgtc actgaagaag aaagatttgt acaaagggta
     3421 gtggactttt tgcaaattca ttgtcaccaa tccatcgttg ccgaaccctt gtctgaagct
     3481 caaactttgg agcaccgtca ggggcaaatt cattactgcc aacaacaaca gaaaaatgat
     3541 aaaacccgtc gggtactgga aaaagctttt ggggaagctt gggcggaacg gtatatgagc
     3601 caagtcttat ttgatgttat ccaataatct agaccaggca tcaaataaaa cgaaaggctc
     3661 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     3721 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatagcacct gaagtcagcc
     3781 ccatacgata taagttgtta ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     3841 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     3901 ggagtccaag cgagctctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     3961 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     4021 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     4081 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     4141 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     4201 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     4261 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     4321 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     4381 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     4441 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     4501 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     4561 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     4621 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4681 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     4741 tcctcatcct gtctcttgat cagatcatga tcccctgcgc catcagatcc ttggcggcaa
     4801 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     4861 caattccgac gtcggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgcattca
     4921 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     4981 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     5041 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     5101 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     5161 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tg
//
