LOCUS       sequence-146322-        4981 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 4981)
  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 4981)
  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-morange
FEATURES             Location/Qualifiers
     source          1..4981
                     /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"
     CDS             2538..2555
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2565..3275
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2978..2997
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3375..3392)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3375..3391)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3384..3406)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3552..4385)
                     /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      4399..4750
                     /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(4812..4831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4931..4953
                     /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 caagaaggat ggggagccac catcaccatc accatggcag atctatggtg agcaagggcg
     2581 aggagaataa catggccatc atcaaggagt tcatgcgctt caaggtgcgc atggagggct
     2641 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggct
     2701 ttcagaccgt taagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatct
     2761 tgtcccctca gttcacctac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     2821 actacctcaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2881 acggcggcgt ggtgaccgtg actcaggact cctccctgca ggacggcgag ttcatctaca
     2941 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     3001 tgggcatgga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgagg
     3061 acaagctcag gctgaagctg aaggacggcg gccactacac ctccgaggtc aagaccacct
     3121 acaaggccaa gaagcccgtg cagttgcccg gcgcctacat cgtcgacatc aagttggaca
     3181 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     3241 actccaccgg cggcatggac gagctgtaca agtaacagta cgaacgcgcc gagggccgcc
     3301 actccaccgg cggcatggac gagctgtaca agtaaggatc ctctagagtc gacctgcagg
     3361 catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3421 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3481 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga aaggcaggcc gggccgtggt
     3541 ggccacggcc tctaggccag atccagcggc atctgggtta gtcgagcgcg ggccgcttcc
     3601 catgtctcac cagggcgagc ctgtttcgcg atctcagcat ctgaaatctt cccggccttg
     3661 cgcttcgctg gggccttacc caccgccttg gcgggcttct tcggtccaaa actgaacaac
     3721 agatgtgtga ccttgcgccc ggtctttcgc tgcgcccact ccacctgtag cgggctgtgc
     3781 tcgttgatct gcgtcacggc tggatcaagc actcgcaact tgaagtcctt gatcgaggga
     3841 taccggcctt ccagttgaaa ccactttcgc agctggtcaa tttctatttc gcgctggccg
     3901 atgctgtccc attgcatgag cagctcgtaa agcctgatcg cgtgggtgct gtccatcttg
     3961 gccacgtcag ccaaggcgta tttggtgaac tgtttggtga gttccgtcag gtacggcagc
     4021 atgtctttgg tgaacctgag ttctacacgg ccctcaccct cccggtagat gattgtttgc
     4081 acccagccgg taatcatcac actcggtctt ttccccttgc cattgggctc ttgggttaac
     4141 cggacttccc gccgtttcag gcgcagggcc gcttctttga gctggttgta ggaagattcg
     4201 atagggacac ccgccatcgt cgctatgtcc tccgccgtca ctgaatacat cacttcatcg
     4261 gtgacaggct cgctcctctt cacctggcta atacaggcca gaacgatccg ctgttcctga
     4321 acactgaggc gatacgcggc ctcgaccagg gcattgcttt tgtaaaccat tgggggtgag
     4381 gccacgttcg acattccttg tgtataaggg gacactgtat ctgcgtccca caatacaaca
     4441 aatccgtccc tttacaacaa caaatccgtc ccttcttaac aacaaatccg tcccttaatg
     4501 gcaacaaatc cgtccctttt taaactctac aggccacgga ttacgtggcc tgtagacgtc
     4561 ctaaaaggtt taaaagggaa aaggaagaaa agggtggaaa cgcaaaaaac gcaccactac
     4621 gtggccccgt tggggccgca tttgtgcccc tgaaggggcg ggggaggcgt ctgggcaatc
     4681 cccgttttac cagtccccta tcgccgcctg agagggcgca ggaagcgagt aatcagggta
     4741 tcgaggcgga ttcacccttg gcgtccaacc agcggcacca gcggcgcctg agaggtatgg
     4801 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4861 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4921 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4981 a
//
