LOCUS       sequence-276224-        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"
     regulatory      1100..1109
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1106..1762
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     LTR             1955..2545
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2570..2586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2570..2586)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2583..2605)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2594..2610
                     /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(2618..2624,2625..2642,2643..2648))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2663..2684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2801..2818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2972..3560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3052..3071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3731..4522,4523..4591))
                     /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     4354..4373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4592..4696)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4764..4782
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4820..4842)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4942..4961
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5155..5177
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5169..5186
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5170..5186
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt
       61 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt
      121 aagcagttcc tgccccggct cagggccaag aacagttgga acagcagaat atgggccaaa
      181 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
      241 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
      301 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
      361 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
      421 cagtcctccg atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt
      481 ttgcatccga atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc
      541 ccacctcggg ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga
      601 ccaccgaccc ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc
      661 tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttggctaac
      721 tagatctgta tctggcggtc ccgcggaaga actgacgagt tcgtattccc ggccgcagcc
      781 cctgggagac gtcccagcgg cctcgggggc ccgttttgtg gcccattctg tatcagttaa
      841 cctacccgag tcggactttt tggagctccg ccactgtccg aggggtacgt ggctttgttg
      901 ggggacgaga gacagagaca cttcccgccc ccgtctgaat ttttgctttc ggttttacgc
      961 cgaaaccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact
     1021 gtgtttctgt atttgtctga aaattagctc gacaaagtta agtaatagtc cctctctcca
     1081 agctcactta caggcggccg ccaccatggt gagtgtgatt aaaccagaga tgaagatgag
     1141 gtactacatg gacggctccg tcaatgggca tgagttcaca attgaaggtg aaggcacagg
     1201 cagaccttac gagggacatc aagagatgac actacgcgtc acaatggccg agggcgggcc
     1261 aatgcctttc gcgtttgact tagtgtcaca cgtgttctgt tacggccaca gagtatttac
     1321 taaatatcca gaagagatac cagactattt caaacaagca tttcctgaag gcctgtcatg
     1381 ggaaaggtcg ttggagttcg aagatggtgg gtccgcttca gtcagtgcgc atataagcct
     1441 tagaggaaac accttctacc acaaatccaa atttactggg gttaactttc ctgccgatgg
     1501 tcctatcatg caaaaccaaa gtgttgattg ggagccatca accgagaaaa ttactgccag
     1561 cgacggagtt ctgaagggtg atgttacgat gtacctaaaa cttgaaggag gcggcaatca
     1621 caaatgccaa atgaagacta cttacaaggc ggcaaaagag attcttgaaa tgccaggaga
     1681 ccattacatc ggccatcgcc tcgtcaggaa aaccgaaggc aacattactg agcaggtaga
     1741 agatgcagta gctcattcct aaggatccaa gcttaggcct gctcgctttc ttgctgtccc
     1801 atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga
     1861 agggccttga gcatctggat tctgcctagc gctaagctta acacgagcca tagatagaat
     1921 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     1981 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     2041 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     2101 gtggtaagca gttcctgccc cggctcaggg ccaagaacag ttggaacagc agaatatggg
     2161 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     2221 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     2281 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     2341 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     2401 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgttctcaat aaaccctctt
     2461 gcagttgcat ccgactcgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     2521 ctgcccacct cgggggtctt tcattctcga gcagcttggc gtaatcatgg tcatagctgt
     2581 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2641 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2701 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2761 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2821 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2881 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2941 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3001 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3301 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3361 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3421 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3481 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3661 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3721 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3781 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3841 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3901 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3961 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4021 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4081 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4141 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4201 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4261 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4321 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4381 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4441 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4501 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4561 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4621 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4681 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4741 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     4801 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4861 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4921 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     4981 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     5041 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     5101 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     5161 cacgacgttg taaaacgacg gccagtgaat tagtact
//
