LOCUS       sequence-149778-        4927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad range bacterial fluorescence labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4927)
  AUTHORS   Barbier M, Damron FH
  TITLE     Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling.
  JOURNAL   PLoS One. 2016 Mar 3;11(3):e0146827. doi: 
            10.1371/journal.pone.0146827. eCollection 2016.
  PUBMED    26937640
REFERENCE   2  (bases 1 to 4927)
  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: "PLoS One.";
            date: "2016-03-3"; volume: "11(3):e0146827. doi"; pages: " 
            10.1371/journal.pone.0146827. eCollection 2016"
COMMENT     Sequence Label: pUCP20T-E2Crimson
FEATURES             Location/Qualifiers
     source          1..4927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            1892..2000
                     /note="incP origin of transfer"
     primer_bind     2250..2267
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2384..2405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2420..2425,2426..2443,2444..2450)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2458..2474
                     /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     2463..2485
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     regulatory      2559..2568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2565..3239
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     2963..2982
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3321..3338)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3321..3337)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3330..3352)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3498..4331)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      4345..4696
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     primer_bind     complement(4758..4777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4877..4899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacata agctagctta tcggccagcc tcgcagagca ggattcccgt
     1921 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc
     1981 ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa
     2041 ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta
     2101 taatgacccc gaagcagggt tatgcagcgg aaagtatacc ttaaggaatc cccatgttct
     2161 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2221 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2281 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2341 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     2401 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     2461 tgagcggata acaatttcac acaggaaaca tgattacgaa ttctttccac tcctccaatg
     2521 ttttagttag aggtgccgat gatgggaatc cactagtcgc caccatggat agcactgaga
     2581 acgtcatcaa gcccttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     2641 agttcgagat cgagggcgtg ggcgagggca agccctacga gggcacccag accgccaagc
     2701 tgcaagtgac caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttct
     2761 tctacggctc caaggcgtac atcaagcacc ccgccgacat ccccgactac ctcaagcagt
     2821 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     2881 ccgtgaccca ggactcctcc ctgcaggacg gcaccctcat ctaccacgtg aagttcatcg
     2941 gcgtgaactt cccctccgac ggccccgtaa tgcagaagaa gactctgggc tgggagccct
     3001 ccactgagcg caactacccc cgcgacggcg tgctgaaggg cgagaaccac atggcgctga
     3061 agctgaaggg cggcggccac tacctgtgtg agttcaagtc catctacatg gccaagaagc
     3121 ccgtgaagct gcccggctac cactacgtgg actacaagct cgacatcacc tcccacaacg
     3181 aggactacac cgtggtggag cagtacgagc gcgccgaggc ccgccaccac ctgttccaga
     3241 ctcacggtat ggacgaattg tacaagcacg acgaattgta aggatcctct agagtcgacc
     3301 tgcaggcatg caagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3361 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3421 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaaagg caggccgggc
     3481 cgtggtggcc acggcctcta ggccagatcc agcggcatct gggttagtcg agcgcgggcc
     3541 gcttcccatg tctcaccagg gcgagcctgt ttcgcgatct cagcatctga aatcttcccg
     3601 gccttgcgct tcgctggggc cttacccacc gccttggcgg gcttcttcgg tccaaaactg
     3661 aacaacagat gtgtgacctt gcgcccggtc tttcgctgcg cccactccac ctgtagcggg
     3721 ctgtgctcgt tgatctgcgt cacggctgga tcaagcactc gcaacttgaa gtccttgatc
     3781 gagggatacc ggccttccag ttgaaaccac tttcgcagct ggtcaatttc tatttcgcgc
     3841 tggccgatgc tgtcccattg catgagcagc tcgtaaagcc tgatcgcgtg ggtgctgtcc
     3901 atcttggcca cgtcagccaa ggcgtatttg gtgaactgtt tggtgagttc cgtcaggtac
     3961 ggcagcatgt ctttggtgaa cctgagttct acacggccct caccctcccg gtagatgatt
     4021 gtttgcaccc agccggtaat catcacactc ggtcttttcc ccttgccatt gggctcttgg
     4081 gttaaccgga cttcccgccg tttcaggcgc agggccgctt ctttgagctg gttgtaggaa
     4141 gattcgatag ggacacccgc catcgtcgct atgtcctccg ccgtcactga atacatcact
     4201 tcatcggtga caggctcgct cctcttcacc tggctaatac aggccagaac gatccgctgt
     4261 tcctgaacac tgaggcgata cgcggcctcg accagggcat tgcttttgta aaccattggg
     4321 ggtgaggcca cgttcgacat tccttgtgta taaggggaca ctgtatctgc gtcccacaat
     4381 acaacaaatc cgtcccttta caacaacaaa tccgtccctt cttaacaaca aatccgtccc
     4441 ttaatggcaa caaatccgtc cctttttaaa ctctacaggc cacggattac gtggcctgta
     4501 gacgtcctaa aaggtttaaa agggaaaagg aagaaaaggg tggaaacgca aaaaacgcac
     4561 cactacgtgg ccccgttggg gccgcatttg tgcccctgaa ggggcggggg aggcgtctgg
     4621 gcaatccccg ttttaccagt cccctatcgc cgcctgagag ggcgcaggaa gcgagtaatc
     4681 agggtatcga ggcggattca cccttggcgt ccaaccagcg gcaccagcgg cgcctgagag
     4741 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4801 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4861 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4921 cgcgcga
//
