LOCUS       sequence-146309-        5045 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 5045)
  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 5045)
  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-ecfp
FEATURES             Location/Qualifiers
     source          1..5045
                     /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"
     primer_bind     2482..2498
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2482..2498
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             2538..2549
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2556..2558,2559..2561,2562..3275)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2601..2622)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2862..2881)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3209..3230
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3298..3392
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3439..3456)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3439..3455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3448..3470)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3616..4449)
                     /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      4463..4814
                     /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(4876..4895)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4995..5017
                     /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 gctatgacca tgattacgaa ttcggcgcgc
     2521 caagaaggtc tagaattaaa gaggagaaat taagcatgct gagcaagggc gaggagctgt
     2581 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2641 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2701 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctggggcg
     2761 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2821 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2881 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2941 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacatcagcc
     3001 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     3061 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3121 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     3181 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3241 ggatcactct cggcatggac gagctgtaca agtaataagc ttaattagct gagcttggac
     3301 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
     3361 gttgccgccg ggcgtttttt attggtgaga atccaagctg gatcctctag agtcgacctg
     3421 caggcatgca agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3481 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3541 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaaaggca ggccgggccg
     3601 tggtggccac ggcctctagg ccagatccag cggcatctgg gttagtcgag cgcgggccgc
     3661 ttcccatgtc tcaccagggc gagcctgttt cgcgatctca gcatctgaaa tcttcccggc
     3721 cttgcgcttc gctggggcct tacccaccgc cttggcgggc ttcttcggtc caaaactgaa
     3781 caacagatgt gtgaccttgc gcccggtctt tcgctgcgcc cactccacct gtagcgggct
     3841 gtgctcgttg atctgcgtca cggctggatc aagcactcgc aacttgaagt ccttgatcga
     3901 gggataccgg ccttccagtt gaaaccactt tcgcagctgg tcaatttcta tttcgcgctg
     3961 gccgatgctg tcccattgca tgagcagctc gtaaagcctg atcgcgtggg tgctgtccat
     4021 cttggccacg tcagccaagg cgtatttggt gaactgtttg gtgagttccg tcaggtacgg
     4081 cagcatgtct ttggtgaacc tgagttctac acggccctca ccctcccggt agatgattgt
     4141 ttgcacccag ccggtaatca tcacactcgg tcttttcccc ttgccattgg gctcttgggt
     4201 taaccggact tcccgccgtt tcaggcgcag ggccgcttct ttgagctggt tgtaggaaga
     4261 ttcgataggg acacccgcca tcgtcgctat gtcctccgcc gtcactgaat acatcacttc
     4321 atcggtgaca ggctcgctcc tcttcacctg gctaatacag gccagaacga tccgctgttc
     4381 ctgaacactg aggcgatacg cggcctcgac cagggcattg cttttgtaaa ccattggggg
     4441 tgaggccacg ttcgacattc cttgtgtata aggggacact gtatctgcgt cccacaatac
     4501 aacaaatccg tccctttaca acaacaaatc cgtcccttct taacaacaaa tccgtccctt
     4561 aatggcaaca aatccgtccc tttttaaact ctacaggcca cggattacgt ggcctgtaga
     4621 cgtcctaaaa ggtttaaaag ggaaaaggaa gaaaagggtg gaaacgcaaa aaacgcacca
     4681 ctacgtggcc ccgttggggc cgcatttgtg cccctgaagg ggcgggggag gcgtctgggc
     4741 aatccccgtt ttaccagtcc cctatcgccg cctgagaggg cgcaggaagc gagtaatcag
     4801 ggtatcgagg cggattcacc cttggcgtcc aaccagcggc accagcggcg cctgagaggt
     4861 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4921 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4981 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5041 cgcga
//
