LOCUS       40924_45353        4890 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUCP20T-tSapphire, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4890)
  AUTHORS   Barbier M, Damron FH.
  TITLE     Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling
  JOURNAL   PLoS ONE 11 (3), E0146827 (2016)
  PUBMED    26937640
REFERENCE   2  (bases 1 to 4890)
  AUTHORS   Barbier M, Damron FH.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-OCT-2015) Microbiology, Immunology and Cell Biology, 
            West Virginia University, School of Medicine, One Medical Center 
            Drive, Morgantown, WV 26506, USA
REFERENCE   3  (bases 1 to 4890)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4890)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2016"; volume: "11"; issue: "3"; pages: "E0146827"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-OCT-2015) Microbiology, Immunology and Cell Biology, West 
            Virginia University, School of Medicine, One Medical Center Drive, 
            Morgantown, WV 26506, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            complement(1892..2000)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    2384..2405
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2420..2450
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2458..2474
                     /label=lac operator
                     /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     2482..2498
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2528..3241
                     /codon_start=1
                     /label=mgfp5
                     /note="GFP with folding enhancement mutations"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     primer_bind     complement(3284..3300)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3464..4294)
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      4308..4659
                     /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)"
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 gctatgacat gattacgaat tcggcgcgcc
     2521 aagaaggatg tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt
     2581 agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac
     2641 ttacggtaaa ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc
     2701 aaccttagtc actacttttt cttatggtgt tatggttttt gctagatacc cagatcatat
     2761 gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat
     2821 ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac
     2881 cttagttaat agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg
     2941 tcacaaattg gaatacaact ttaactctca caatgtttac atcatggctg acaaacaaaa
     3001 gaatggtatc aaagctaact tcaaaattag acacaacatt gaagatggtg gtgttcaatt
     3061 agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa
     3121 ccattactta tccattcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat
     3181 ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtattgatg aattgtacaa
     3241 ataaggatcc tctagagtcg acctgcaggc atgcaagctt ggcactggcc gtcgttttac
     3301 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3361 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3421 gcagcctgaa aggcaggccg ggccgtggtg gccacggcct ctaggccaga tccagcggca
     3481 tctgggttag tcgagcgcgg gccgcttccc atgtctcacc agggcgagcc tgtttcgcga
     3541 tctcagcatc tgaaatcttc ccggccttgc gcttcgctgg ggccttaccc accgccttgg
     3601 cgggcttctt cggtccaaaa ctgaacaaca gatgtgtgac cttgcgcccg gtctttcgct
     3661 gcgcccactc cacctgtagc gggctgtgct cgttgatctg cgtcacggct ggatcaagca
     3721 ctcgcaactt gaagtccttg atcgagggat accggccttc cagttgaaac cactttcgca
     3781 gctggtcaat ttctatttcg cgctggccga tgctgtccca ttgcatgagc agctcgtaaa
     3841 gcctgatcgc gtgggtgctg tccatcttgg ccacgtcagc caaggcgtat ttggtgaact
     3901 gtttggtgag ttccgtcagg tacggcagca tgtctttggt gaacctgagt tctacacggc
     3961 cctcaccctc ccggtagatg attgtttgca cccagccggt aatcatcaca ctcggtcttt
     4021 tccccttgcc attgggctct tgggttaacc ggacttcccg ccgtttcagg cgcagggccg
     4081 cttctttgag ctggttgtag gaagattcga tagggacacc cgccatcgtc gctatgtcct
     4141 ccgccgtcac tgaatacatc acttcatcgg tgacaggctc gctcctcttc acctggctaa
     4201 tacaggccag aacgatccgc tgttcctgaa cactgaggcg atacgcggcc tcgaccaggg
     4261 cattgctttt gtaaaccatt gggggtgagg ccacgttcga cattccttgt gtataagggg
     4321 acactgtatc tgcgtcccac aatacaacaa atccgtccct ttacaacaac aaatccgtcc
     4381 cttcttaaca acaaatccgt cccttaatgg caacaaatcc gtcccttttt aaactctaca
     4441 ggccacggat tacgtggcct gtagacgtcc taaaaggttt aaaagggaaa aggaagaaaa
     4501 gggtggaaac gcaaaaaacg caccactacg tggccccgtt ggggccgcat ttgtgcccct
     4561 gaaggggcgg gggaggcgtc tgggcaatcc ccgttttacc agtcccctat cgccgcctga
     4621 gagggcgcag gaagcgagta atcagggtat cgaggcggat tcacccttgg cgtccaacca
     4681 gcggcaccag cggcgcctga gaggtatggt gcactctcag tacaatctgc tctgatgccg
     4741 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4801 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4861 ggttttcacc gtcatcaccg aaacgcgcga
//
