LOCUS       40924_31160        3439 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMN10, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3439)
  AUTHORS   Jiwaji M, Daly R, Gibriel A, Barkess G, McLean P, Yang J, Pansare K,
            Cumming S, McLauchlan A, Kamola PJ, Bhutta MS, West AG, West KL, 
            Kolch W, Girolami MA, Pitt AR.
  TITLE     Unique reporter-based sensor platforms to monitor signalling in 
            cells
  JOURNAL   PLoS ONE 7 (11), E50521 (2012)
  PUBMED    23209767
REFERENCE   2  (bases 1 to 3439)
  AUTHORS   Jiwaji M.
  TITLE     The Molecular Nose
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3439)
  AUTHORS   Jiwaji M.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-NOV-2009) FBLS, Glasgow University, University Avenue,
            Glasgow G12 8QQ, UK
REFERENCE   4  (bases 1 to 3439)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3439)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2012"; volume: "7"; issue: "11"; pages: "E50521"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (17-NOV-2009) FBLS, Glasgow University, University Avenue, Glasgow 
            G12 8QQ, UK"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           137..176
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     misc_feature    194..316
                     /label=unique reporter sequence
                     /note="unique reporter sequence"
     polyA_signal    complement(350..471)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(890..1478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1652..2509)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2510..2614)
                     /label=AmpR promoter
     rep_origin      2641..3096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    3227..3275
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    3289..3380
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     protein_bind    3406..3439
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ctcgagggaa ttccgagctc ggatatcccg actcgtccaa cgactataaa gagggcaggc
       61 tgtcctctaa gcgtcaccac gacttcaacg tcctgagtac cttctcctca cttactccgt
      121 agctccagct tcaccagatc ttggtggcgt gaaactcccg cacctcttcg gcaagcgcct
      181 tgtagaagcg cgtaagcttg gacttagcgg actgtgtgga cgggaactga gtgagatgag
      241 cgtgcgagag tgccacgcct ggcagatgtc ggtcggagag gaccttgatg tgtgtggctc
      301 taacggtcgg tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
      361 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
      421 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
      481 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
      541 taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc ccttgagagc
      601 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
      661 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
      721 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      781 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      841 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      901 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      961 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1021 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1081 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1141 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1201 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1261 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1321 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1381 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1441 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1501 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1561 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1621 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1681 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1741 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1801 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1861 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1921 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1981 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2041 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2101 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2161 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2221 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2281 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2341 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2401 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2461 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2521 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2581 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2641 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2701 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2761 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2821 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2881 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2941 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3001 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3061 acgcgaattt taacaaaata ttaacgttta caatttccca ttcgccattc aggctgcgca
     3121 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
     3181 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
     3241 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
     3301 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
     3361 aggtgccaga acatttctct atcgataggt acccccgggc cgcggataac ttcgtataat
     3421 gtatgctata cgaagttat
//
