LOCUS       sequence-151520-        5020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad range bacterial fluorescence labeling (ECFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5020)
  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 5020)
  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-eCFP
FEATURES             Location/Qualifiers
     source          1..5020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      72..660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     561..580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            732..840
                     /note="incP origin of transfer"
     primer_bind     1090..1107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1224..1245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1260..1265,1266..1283,1284..1290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1298..1314
                     /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     1303..1325
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1322..1338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1322..1338
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             1377..1388
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1395..1397,1398..1400,1401..2114)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1440..1461)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1701..1720)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2048..2069
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2125..2142)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2125..2141)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2134..2156)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2302..3135)
                     /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      3149..3500
                     /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(3541..3546,3547..3563,3564..3569)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3758..4291
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
       61 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
      241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacata
      721 agctagctta tcggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
      781 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
      841 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
      901 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
      961 tatgcagcgg aaagtatacc ttaaggaatc cccatgttct ttcctgcgtt atcccctgat
     1021 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1081 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     1141 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1201 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     1261 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     1321 acaggaaaca gctatgacat gattacgaat tcggcgcgcc aagaaggtct agaattaaag
     1381 aggagaaatt aagcatgctg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1441 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1501 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1561 tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc
     1621 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1681 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1741 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1801 acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg
     1861 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggca
     1921 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1981 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2041 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2101 agctgtacaa gtaataagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     2161 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     2221 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga aaggcaggcc
     2281 gggccgtggt ggccacggcc tctaggccag atccagcggc atctgggtta gtcgagcgcg
     2341 ggccgcttcc catgtctcac cagggcgagc ctgtttcgcg atctcagcat ctgaaatctt
     2401 cccggccttg cgcttcgctg gggccttacc caccgccttg gcgggcttct tcggtccaaa
     2461 actgaacaac agatgtgtga ccttgcgccc ggtctttcgc tgcgcccact ccacctgtag
     2521 cgggctgtgc tcgttgatct gcgtcacggc tggatcaagc actcgcaact tgaagtcctt
     2581 gatcgaggga taccggcctt ccagttgaaa ccactttcgc agctggtcaa tttctatttc
     2641 gcgctggccg atgctgtccc attgcatgag cagctcgtaa agcctgatcg cgtgggtgct
     2701 gtccatcttg gccacgtcag ccaaggcgta tttggtgaac tgtttggtga gttccgtcag
     2761 gtacggcagc atgtctttgg tgaacctgag ttctacacgg ccctcaccct cccggtagat
     2821 gattgtttgc acccagccgg taatcatcac actcggtctt ttccccttgc cattgggctc
     2881 ttgggttaac cggacttccc gccgtttcag gcgcagggcc gcttctttga gctggttgta
     2941 ggaagattcg atagggacac ccgccatcgt cgctatgtcc tccgccgtca ctgaatacat
     3001 cacttcatcg gtgacaggct cgctcctctt cacctggcta atacaggcca gaacgatccg
     3061 ctgttcctga acactgaggc gatacgcggc ctcgaccagg gcattgcttt tgtaaaccat
     3121 tgggggtgag gccacgttcg acattccttg tgtataaggg gacactgtat ctgcgtccca
     3181 caatacaaca aatccgtccc tttacaacaa caaatccgtc ccttcttaac aacaaatccg
     3241 tcccttaatg gcaacaaatc cgtccctttt taaactctac aggccacgga ttacgtggcc
     3301 tgtagacgtc ctaaaaggtt taaaagggaa aaggaagaaa agggtggaaa cgcaaaaaac
     3361 gcaccactac gtggccccgt tggggccgca tttgtgcccc tgaaggggcg ggggaggcgt
     3421 ctgggcaatc cccgttttac cagtccccta tcgccgcctg agagggcgca ggaagcgagt
     3481 aatcagggta tcgaggcgga ttcacccttg gcgtccaacc agcggcacca gcggcgcgaa
     3541 ttgacataag cctgttcggt tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac
     3601 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     3661 tgttatgact gtttttttgt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     3721 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gcaacgatgt
     3781 tacgcagcag ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg
     3841 cacatgtagg ctcggccctg accaagtcaa atccatgcgg gctgctcttg atcttttcgg
     3901 tcgtgagttc ggagacgtag ccacctactc ccaacatcag ccggactccg attacctcgg
     3961 gaacttgctc cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt
     4021 tggcgctctc gcggcttacg ttctgcccag gtttgagcag ccgcgtagtg agatctatat
     4081 ctatgatctc gcagtctccg gcgagcaccg gaggcagggc attgccaccg cgctcatcaa
     4141 tctcctcaag catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta
     4201 cggtgacgat cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca
     4261 ctttgatatc gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc
     4321 ggccgcggag ttgttcggta aattgtcaca acgccgcggc caattcgact caccagtcac
     4381 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4441 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4501 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4561 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4621 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4681 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4741 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4801 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4861 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4921 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4981 taaaaggatc taggtgaaga tcctttttga taatctcatg
//
