LOCUS       sequence-206122-        5276 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 5276)
  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 5276)
  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: pUCP30T-E2Crimson
FEATURES             Location/Qualifiers
     source          1..5276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     regulatory      193..202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             199..873
                     /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     597..616
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1062..1078)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1071..1093)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1239..2072)
                     /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      2086..2437
                     /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)"
     promoter        join(2478..2483,2484..2500,2501..2506)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2695..3228
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      4029..4617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4518..4537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            4687..4795
                     /note="incP origin of transfer"
     primer_bind     5045..5062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5179..5200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5215..5220,5221..5238,5239..5245)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5253..5269
                     /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     join(5258..5276,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat ggaattctgt aaaggtaata ttaccacgaa aagggcgtac
       61 aggaccacgg agagtggtac cagcagcttg ccagcgctta gtctgttgaa aaacatcctt
      121 cttggcgcgc cgaattcttt ccactcctcc aatgttttag ttagaggtgc cgatgatggg
      181 gatccactag tcgccaccat ggatagcact gagaacgtca tcaagccctt catgcgcttc
      241 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgtgggcgag
      301 ggcaagccct acgagggcac ccagaccgcc aagctgcaag tgaccaaggg cggccccctg
      361 cccttcgcct gggacatcct gtccccccag ttcttctacg gctccaaggc gtacatcaag
      421 caccccgccg acatccccga ctacctcaag cagtccttcc ccgagggctt caagtgggag
      481 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      541 gacggcaccc tcatctacca cgtgaagttc atcggcgtga acttcccctc cgacggcccc
      601 gtaatgcaga agaagactct gggctgggag ccctccactg agcgcaacta cccccgcgac
      661 ggcgtgctga agggcgagaa ccacatggcg ctgaagctga agggcggcgg ccactacctg
      721 tgtgagttca agtccatcta catggccaag aagcccgtga agctgcccgg ctaccactac
      781 gtggactaca agctcgacat cacctcccac aacgaggact acaccgtggt ggagcagtac
      841 gagcgcgccg aggcccgcca ccacctgttc cagactcacg gtatggacga attgtacaag
      901 cacgacgaat tgtaaggatc ccactggcct ttactgaggt cctggggatt gaaattcaga
      961 tccgcatgct tgtctggtaa gtactttacc caccgaacca tcttcccaac cctttagctt
     1021 cctcttggct tatgaaaagc caagacggcc agtgggatcc cactggccgt cgttttacaa
     1081 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1141 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1201 agcctgaaag gcaggccggg ccgtggtggc cacggcctct aggccagatc cagcggcatc
     1261 tgggttagtc gagcgcgggc cgcttcccat gtctcaccag ggcgagcctg tttcgcgatc
     1321 tcagcatctg aaatcttccc ggccttgcgc ttcgctgggg ccttacccac cgccttggcg
     1381 ggcttcttcg gtccaaaact gaacaacaga tgtgtgacct tgcgcccggt ctttcgctgc
     1441 gcccactcca cctgtagcgg gctgtgctcg ttgatctgcg tcacggctgg atcaagcact
     1501 cgcaacttga agtccttgat cgagggatac cggccttcca gttgaaacca ctttcgcagc
     1561 tggtcaattt ctatttcgcg ctggccgatg ctgtcccatt gcatgagcag ctcgtaaagc
     1621 ctgatcgcgt gggtgctgtc catcttggcc acgtcagcca aggcgtattt ggtgaactgt
     1681 ttggtgagtt ccgtcaggta cggcagcatg tctttggtga acctgagttc tacacggccc
     1741 tcaccctccc ggtagatgat tgtttgcacc cagccggtaa tcatcacact cggtcttttc
     1801 cccttgccat tgggctcttg ggttaaccgg acttcccgcc gtttcaggcg cagggccgct
     1861 tctttgagct ggttgtagga agattcgata gggacacccg ccatcgtcgc tatgtcctcc
     1921 gccgtcactg aatacatcac ttcatcggtg acaggctcgc tcctcttcac ctggctaata
     1981 caggccagaa cgatccgctg ttcctgaaca ctgaggcgat acgcggcctc gaccagggca
     2041 ttgcttttgt aaaccattgg gggtgaggcc acgttcgaca ttccttgtgt ataaggggac
     2101 actgtatctg cgtcccacaa tacaacaaat ccgtcccttt acaacaacaa atccgtccct
     2161 tcttaacaac aaatccgtcc cttaatggca acaaatccgt ccctttttaa actctacagg
     2221 ccacggatta cgtggcctgt agacgtccta aaaggtttaa aagggaaaag gaagaaaagg
     2281 gtggaaacgc aaaaaacgca ccactacgtg gccccgttgg ggccgcattt gtgcccctga
     2341 aggggcgggg gaggcgtctg ggcaatcccc gttttaccag tcccctatcg ccgcctgaga
     2401 gggcgcagga agcgagtaat cagggtatcg aggcggattc acccttggcg tccaaccagc
     2461 ggcaccagcg gcgcgaattg acataagcct gttcggttcg taaactgtaa tgcaagtagc
     2521 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     2581 tggcggtttt catggcttgt tatgactgtt tttttgtaca gtctatgcct cgggcatcca
     2641 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     2701 cgcagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggtggctca
     2761 agtatgggca tcattcgcac atgtaggctc ggccctgacc aagtcaaatc catgcgggct
     2821 gctcttgatc ttttcggtcg tgagttcgga gacgtagcca cctactccca acatcagccg
     2881 gactccgatt acctcgggaa cttgctccgt agtaagacat tcatcgcgct tgctgccttc
     2941 gaccaagaag cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg
     3001 cgtagtgaga tctatatcta tgatctcgca gtctccggcg agcaccggag gcagggcatt
     3061 gccaccgcgc tcatcaatct cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc
     3121 tacgtgcaag cagattacgg tgacgatccc gcagtggctc tctatacaaa gttgggcata
     3181 cgggaagaag tgatgcactt tgatatcgac ccaagtaccg ccacctaaca attcgttcaa
     3241 gccgagatcg gcttcccggc cgcggagttg ttcggtaaat tgtcacaacg ccgcggccaa
     3301 ttcgactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3361 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3421 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3481 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3541 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3601 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3661 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3721 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3781 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3841 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3901 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3961 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4021 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4081 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4141 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4201 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4261 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4321 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4381 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4441 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4501 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4561 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4621 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacaagcta
     4681 gcttatcggc cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg
     4741 gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg
     4801 acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg
     4861 tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc
     4921 agcggaaagt ataccttaag gaatccccat gttctttcct gcgttatccc ctgattctgt
     4981 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5041 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     5101 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5161 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     5221 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcaca
//
