LOCUS       sequence-304891-        5197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMP71 expression vector containing mKO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5197)
  AUTHORS   Bresser K, Dijkgraaf FE, Pritchard CEJ, Huijbers IJ, Song JY, Rohr 
            JC, Scheeren FA, Schumacher TN
  TITLE     A mouse model that is immunologically tolerant to reporter and 
            modifier proteins.
  JOURNAL   Commun Biol. 2020 May 29;3(1):273. doi: 10.1038/s42003-020-0979-0.
  PUBMED    32472011
REFERENCE   2  (bases 1 to 5197)
  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: "Commun 
            Biol."; date: "2020-05-29"; volume: "3(1):273. doi"; pages: " 
            10.1038/s42003-020-0979-0"
COMMENT     Sequence Label: pMP71-mKO2
FEATURES             Location/Qualifiers
     source          1..5197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1134..1790
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     LTR             1983..2573
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2598..2614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2598..2614)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2611..2633)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2622..2638
                     /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)."
     promoter        complement(join(2646..2652,2653..2670,2671..2676))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2691..2712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2829..2846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3000..3588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3080..3099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3759..4550,4551..4619))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4382..4401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4620..4724)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4792..4810
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4848..4870)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4970..4989
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5183..5197,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5197,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttagtactct agcttaagta acgccatttt gcaaggcatg
       61 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
      121 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
      181 cagttggaac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
      241 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
      301 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggactcg ctgatccttg
      541 ggagggtctc ctcagattga ttgactgccc acctcggggg tctttcattt ggaggttcca
      601 ccgagatttg gagacccctg cccagggacc accgaccccc ccgccgggag gtaagctggc
      661 cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt
      721 gcgcctgcgt ctgtactagt tggctaacta gatctgtatc tggcggtccc gcggaagaac
      781 tgacgagttc gtattcccgg ccgcagcccc tgggagacgt cccagcggcc tcgggggccc
      841 gttttgtggc ccattctgta tcagttaacc tacccgagtc ggactttttg gagctccgcc
      901 actgtccgag gggtacgtgg ctttgttggg ggacgagaga cagagacact tcccgccccc
      961 gtctgaattt ttgctttcgg ttttacgccg aaaccgcgcc gcgcgtcttg tctgctgcag
     1021 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagctcga
     1081 caaagttaag taatagtccc tctctccaag ctcacttaca ggcggccgcc accatggtga
     1141 gtgtgattaa accagagatg aagatgaggt actacatgga cggctccgtc aatgggcatg
     1201 agttcacaat tgaaggtgaa ggcacaggca gaccttacga gggacatcaa gagatgacac
     1261 tacgcgtcac aatggccgag ggcgggccaa tgcctttcgc gtttgactta gtgtcacacg
     1321 tgttctgtta cggccacaga gtatttacta aatatccaga agagatacca gactatttca
     1381 aacaagcatt tcctgaaggc ctgtcatggg aaaggtcgtt ggagttcgaa gatggtgggt
     1441 ccgcttcagt cagtgcgcat ataagcctta gaggaaacac cttctaccac aaatccaaat
     1501 ttactggggt taactttcct gccgatggtc ctatcatgca aaaccaaagt gttgattggg
     1561 agccatcaac cgagaaaatt actgccagcg acggagttct gaagggtgat gttacgatgt
     1621 acctaaaact tgaaggaggc ggcaatcaca aatgccaaat gaagactact tacaaggcgg
     1681 caaaagagat tcttgaaatg ccaggagacc attacatcgg ccatcgcctc gtcaggaaaa
     1741 ccgaaggcaa cattactgag caggtagaag atgcagtagc tcattcctaa ggatccaagc
     1801 ttaggcctgc tcgctttctt gctgtcccat ttctattaaa ggttcctttg ttccctaagt
     1861 ccaactacta aactggggga tattatgaag ggccttgagc atctggattc tgcctagcgc
     1921 taagcttaac acgagccata gatagaataa aagattttat ttagtctcca gaaaaagggg
     1981 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     2041 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     2101 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     2161 aagaacagtt ggaacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     2221 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct
     2281 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     2341 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     2401 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg
     2461 ggtacccgtg ttctcaataa accctcttgc agttgcatcc gactcgtggt ctcgctgttc
     2521 cttgggaggg tctcctctga gtgattgact gcccacctcg ggggtctttc attctcgagc
     2581 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2641 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2701 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2761 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2821 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2881 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2941 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3001 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3061 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3121 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3181 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3241 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3301 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3361 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3421 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3481 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3541 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3601 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3661 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3721 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3781 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3841 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3901 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3961 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4021 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4081 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4141 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4201 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4261 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4321 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4381 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4441 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4501 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4561 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4621 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4681 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4741 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4801 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4861 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4921 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4981 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     5041 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     5101 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     5161 agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
