LOCUS       sequence-146320-        4831 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 4831)
  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 4831)
  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-mko1
FEATURES             Location/Qualifiers
     source          1..4831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      232..583
                     /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)"
     CDS             597..1430
                     /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"
     primer_bind     1576..1598
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1590..1607
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1591..1607
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1647..2303)
                     /codon_start=1
                     /product="Kusabira-Orange fluorescent protein (Karasawa et 
                     al., 2004)"
                     /label=Kusabira-Orange
                     /translation="MVSVIKPEMKMKYFMDGSVNGHEFTVEGEGTGKPYEGHQEMTLRV
                     TMAKGGPMPFSFDLVSHTFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLQFEDGGFA
                     AVSAHISLRGNCFEHKSKFVGVNFPADGPVMQNQSSDWEPSTEKITTCDGVLKGDVTMY
                     LKLAGGGNHKCQFKTTYKAAKKILKMPQSHFIGHRLVRKTEGNITELVEDAVAHC"
     primer_bind     complement(2334..2350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2334..2350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2347..2369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2358..2374
                     /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)."
     promoter        complement(join(2382..2388,2389..2406,2407..2412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2427..2448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2565..2582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            2832..2940
                     /note="incP origin of transfer"
     rep_origin      complement(3012..3600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3092..3111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3771..4562,4563..4631))
                     /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     4394..4413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4632..4736)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4804..4822
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatacctc tcaggcgccg ctggtgccgc tggttggacg ccaagggtga atccgcctcg
      241 ataccctgat tactcgcttc ctgcgccctc tcaggcggcg ataggggact ggtaaaacgg
      301 ggattgccca gacgcctccc ccgccccttc aggggcacaa atgcggcccc aacggggcca
      361 cgtagtggtg cgttttttgc gtttccaccc ttttcttcct tttccctttt aaacctttta
      421 ggacgtctac aggccacgta atccgtggcc tgtagagttt aaaaagggac ggatttgttg
      481 ccattaaggg acggatttgt tgttaagaag ggacggattt gttgttgtaa agggacggat
      541 ttgttgtatt gtgggacgca gatacagtgt ccccttatac acaaggaatg tcgaacgtgg
      601 cctcaccccc aatggtttac aaaagcaatg ccctggtcga ggccgcgtat cgcctcagtg
      661 ttcaggaaca gcggatcgtt ctggcctgta ttagccaggt gaagaggagc gagcctgtca
      721 ccgatgaagt gatgtattca gtgacggcgg aggacatagc gacgatggcg ggtgtcccta
      781 tcgaatcttc ctacaaccag ctcaaagaag cggccctgcg cctgaaacgg cgggaagtcc
      841 ggttaaccca agagcccaat ggcaagggga aaagaccgag tgtgatgatt accggctggg
      901 tgcaaacaat catctaccgg gagggtgagg gccgtgtaga actcaggttc accaaagaca
      961 tgctgccgta cctgacggaa ctcaccaaac agttcaccaa atacgccttg gctgacgtgg
     1021 ccaagatgga cagcacccac gcgatcaggc tttacgagct gctcatgcaa tgggacagca
     1081 tcggccagcg cgaaatagaa attgaccagc tgcgaaagtg gtttcaactg gaaggccggt
     1141 atccctcgat caaggacttc aagttgcgag tgcttgatcc agccgtgacg cagatcaacg
     1201 agcacagccc gctacaggtg gagtgggcgc agcgaaagac cgggcgcaag gtcacacatc
     1261 tgttgttcag ttttggaccg aagaagcccg ccaaggcggt gggtaaggcc ccagcgaagc
     1321 gcaaggccgg gaagatttca gatgctgaga tcgcgaaaca ggctcgccct ggtgagacat
     1381 gggaagcggc ccgcgctcga ctaacccaga tgccgctgga tctggcctag aggccgtggc
     1441 caccacggcc cggcctgcct ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1501 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1561 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat
     1621 gcctgcaggt cgactctaga ggatcctcag caatgagcta ctgcatcttc taccagctca
     1681 gtaatgttgc cttcggtttt cctgacgagg cgatgcccga tgaaatggct ttgtggcatt
     1741 ttaagaatct tttttgccgc cttgtaagta gtcttgaatt ggcatttgtg attgccgcct
     1801 cccgcaagtt ttaggtacat cgtaacatca cccttcagaa ctccgtcgca ggtagtaatt
     1861 ttctcggttg atggctccca atcagaactt tggttttgca tcacaggacc atcggcagga
     1921 aagttaaccc caacaaattt ggatttgtgc tcgaagcagt ttcctctaag gcttatatgc
     1981 gcactgactg cagcgaaccc accatcttcg aactgcaacg acctttccca tgacaggcct
     2041 tcaggaaatg cttgtttgaa atagtctggt atctcttctg gatatttagt aaaaggtctg
     2101 tggccgtaac agaacgtgtg tgacactaag tcaaacgaga aaggcattgg cccgcccttg
     2161 gccattgtga cgcgtagtgt catctcttga tgtccctcgt aaggtttgcc tgtgccttca
     2221 ccttcaactg tgaactcatg cccattgacg gagccgtcca tgaagtactt catcttcatc
     2281 tctggtttaa tcacactcac catccttctt ggcgcgccga attcgtaatc atggtcatag
     2341 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2401 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2461 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2521 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2581 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2641 ttatccacag aatcagggga taacgcagga aagaacatgg ggattcctta aggtatactt
     2701 tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt
     2761 ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa
     2821 cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc
     2881 actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg
     2941 ataagctagc ttatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3001 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3061 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3121 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3181 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3241 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3301 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3361 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3421 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     3481 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3541 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3601 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3661 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3721 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3781 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3841 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3901 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3961 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4021 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4081 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4141 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4201 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4261 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4321 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4381 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4441 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4501 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4561 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4621 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4681 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4741 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4801 aaaaataggc gtatcacgag gccctttcgt c
//
