LOCUS       40924_45383        5050 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUCP30T-mOrange, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5050)
  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 5050)
  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 5050)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5050)
  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..5050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      72..660
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            complement(732..840)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    1224..1245
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1260..1290
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1298..1314
                     /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     1322..1338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1378..1395
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             1405..2112
                     /codon_start=1
                     /label=LSSmOrange
                     /note="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2155..2171)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2335..3165)
                     /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      3179..3530
                     /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        3571..3599
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3788..4318
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /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 gctatgacca tgattacgaa ttcggcgcgc caagaaggat ggggagccac
     1381 catcaccatc accatggcag atctatggtg agcaagggcg aggagaataa catggccatc
     1441 atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg ccacgagttc
     1501 gagatcgagg gcgagggcga gggccgcccc tacgagggct ttcagaccgt taagctgaag
     1561 gtgaccaagg gtggccccct gcccttcgcc tgggacatct tgtcccctca gttcacctac
     1621 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacctcaa gctgtccttc
     1681 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1741 actcaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1801 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggcatgga ggcctcctcc
     1861 gagcggatgt accccgagga cggcgccctg aagggcgagg acaagctcag gctgaagctg
     1921 aaggacggcg gccactacac ctccgaggtc aagaccacct acaaggccaa gaagcccgtg
     1981 cagttgcccg gcgcctacat cgtcgacatc aagttggaca tcacctccca caacgaggac
     2041 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2101 gagctgtaca agtaaggatc ctctagagtc gacctgcagg catgcaagct tggcactggc
     2161 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     2221 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     2281 ccaacagttg cgcagcctga aaggcaggcc gggccgtggt ggccacggcc tctaggccag
     2341 atccagcggc atctgggtta gtcgagcgcg ggccgcttcc catgtctcac cagggcgagc
     2401 ctgtttcgcg atctcagcat ctgaaatctt cccggccttg cgcttcgctg gggccttacc
     2461 caccgccttg gcgggcttct tcggtccaaa actgaacaac agatgtgtga ccttgcgccc
     2521 ggtctttcgc tgcgcccact ccacctgtag cgggctgtgc tcgttgatct gcgtcacggc
     2581 tggatcaagc actcgcaact tgaagtcctt gatcgaggga taccggcctt ccagttgaaa
     2641 ccactttcgc agctggtcaa tttctatttc gcgctggccg atgctgtccc attgcatgag
     2701 cagctcgtaa agcctgatcg cgtgggtgct gtccatcttg gccacgtcag ccaaggcgta
     2761 tttggtgaac tgtttggtga gttccgtcag gtacggcagc atgtctttgg tgaacctgag
     2821 ttctacacgg ccctcaccct cccggtagat gattgtttgc acccagccgg taatcatcac
     2881 actcggtctt ttccccttgc cattgggctc ttgggttaac cggacttccc gccgtttcag
     2941 gcgcagggcc gcttctttga gctggttgta ggaagattcg atagggacac ccgccatcgt
     3001 cgctatgtcc tccgccgtca ctgaatacat cacttcatcg gtgacaggct cgctcctctt
     3061 cacctggcta atacaggcca gaacgatccg ctgttcctga acactgaggc gatacgcggc
     3121 ctcgaccagg gcattgcttt tgtaaaccat tgggggtgag gccacgttcg acattccttg
     3181 tgtataaggg gacactgtat ctgcgtccca caatacaaca aatccgtccc tttacaacaa
     3241 caaatccgtc ccttcttaac aacaaatccg tcccttaatg gcaacaaatc cgtccctttt
     3301 taaactctac aggccacgga ttacgtggcc tgtagacgtc ctaaaaggtt taaaagggaa
     3361 aaggaagaaa agggtggaaa cgcaaaaaac gcaccactac gtggccccgt tggggccgca
     3421 tttgtgcccc tgaaggggcg ggggaggcgt ctgggcaatc cccgttttac cagtccccta
     3481 tcgccgcctg agagggcgca ggaagcgagt aatcagggta tcgaggcgga ttcacccttg
     3541 gcgtccaacc agcggcacca gcggcgcgaa ttgacataag cctgttcggt tcgtaaactg
     3601 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     3661 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt acagtctatg
     3721 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     3781 agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     3841 gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa
     3901 atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag ccacctactc
     3961 ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc
     4021 gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccag
     4081 gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg gcgagcaccg
     4141 gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg
     4201 tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac
     4261 aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta
     4321 acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta aattgtcaca
     4381 acgccgcggc caattcgact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4441 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4501 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4561 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4621 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4681 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4741 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4801 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4861 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4921 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4981 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5041 taatctcatg
//
