LOCUS       sequence-394374-        4888 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 4888)
  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 4888)
  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-eyfp
FEATURES             Location/Qualifiers
     source          1..4888
                     /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            1890..1998
                     /note="incP origin of transfer"
     primer_bind     2248..2265
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2382..2403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2418..2423,2424..2441,2442..2448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2456..2472
                     /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     2461..2483
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2480..2496
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2480..2496
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             2535..2546
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2553..2555,2556..2558,2559..3272)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2598..2619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2859..2878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3206..3227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3283..3300)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3283..3299)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3292..3314)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3460..4293)
                     /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      4307..4658
                     /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(4719..4738)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4838..4860
                     /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 aatactgttc
     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 tgctcacaag ctagcttatc ggccagcctc gcagagcagg attcccgttg
     1921 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     1981 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     2041 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     2101 atgaccccga agcagggtta tgcagcggaa agtatacctt aaggaatccc catgttcttt
     2161 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2221 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2281 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2341 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2401 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2461 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattc ggcgcgccaa
     2521 gaaggtctag aattaaagag gagaaattaa gcatgctgag caagggcgag gagctgttca
     2581 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     2641 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     2701 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc
     2761 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2821 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2881 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2941 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3001 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     3061 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3121 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca
     3181 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3241 tcactctcgg catggacgag ctgtacaagt aataagcttg gcactggccg tcgttttaca
     3301 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     3361 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3421 cagcctgaaa ggcaggccgg gccgtggtgg ccacggcctc taggccagat ccagcggcat
     3481 ctgggttagt cgagcgcggg ccgcttccca tgtctcacca gggcgagcct gtttcgcgat
     3541 ctcagcatct gaaatcttcc cggccttgcg cttcgctggg gccttaccca ccgccttggc
     3601 gggcttcttc ggtccaaaac tgaacaacag atgtgtgacc ttgcgcccgg tctttcgctg
     3661 cgcccactcc acctgtagcg ggctgtgctc gttgatctgc gtcacggctg gatcaagcac
     3721 tcgcaacttg aagtccttga tcgagggata ccggccttcc agttgaaacc actttcgcag
     3781 ctggtcaatt tctatttcgc gctggccgat gctgtcccat tgcatgagca gctcgtaaag
     3841 cctgatcgcg tgggtgctgt ccatcttggc cacgtcagcc aaggcgtatt tggtgaactg
     3901 tttggtgagt tccgtcaggt acggcagcat gtctttggtg aacctgagtt ctacacggcc
     3961 ctcaccctcc cggtagatga ttgtttgcac ccagccggta atcatcacac tcggtctttt
     4021 ccccttgcca ttgggctctt gggttaaccg gacttcccgc cgtttcaggc gcagggccgc
     4081 ttctttgagc tggttgtagg aagattcgat agggacaccc gccatcgtcg ctatgtcctc
     4141 cgccgtcact gaatacatca cttcatcggt gacaggctcg ctcctcttca cctggctaat
     4201 acaggccaga acgatccgct gttcctgaac actgaggcga tacgcggcct cgaccagggc
     4261 attgcttttg taaaccattg ggggtgaggc cacgttcgac attccttgtg tataagggga
     4321 cactgtatct gcgtcccaca atacaacaaa tccgtccctt tacaacaaca aatccgtccc
     4381 ttcttaacaa caaatccgtc ccttaatggc aacaaatccg tcccttttta aactctacag
     4441 gccacggatt acgtggcctg tagacgtcct aaaaggttta aaagggaaaa ggaagaaaag
     4501 ggtggaaacg caaaaaacgc accactacgt ggccccgttg gggccgcatt tgtgcccctg
     4561 aaggggcggg ggaggcgtct gggcaatccc cgttttacca gtcccctatc gccgcctgag
     4621 agggcgcagg aagcgagtaa tcagggtatc gaggcggatt cacccttggc gtccaaccag
     4681 cggcaccagc ggcgcctgag aggatggtgc actctcagta caatctgctc tgatgccgca
     4741 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4801 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4861 ttttcaccgt catcaccgaa acgcgcga
//
