LOCUS       sequence-230490-        7634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMP71 vector expressing Tomato - P2A - CD40L - SrtA - flag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7634)
  AUTHORS   Pasqual G, Chudnovskiy A, Tas JMJ, Agudelo M, Schweitzer LD, Cui A, 
            Hacohen N, Victora GD
  TITLE     Monitoring T cell-dendritic cell interactions in vivo by 
            intercellular enzymatic labelling.
  JOURNAL   Nature. 2018 Jan 25;553(7689):496-500. doi: 10.1038/nature25442. 
            Epub 2018 Jan 17.
  PUBMED    29342141
REFERENCE   2  (bases 1 to 7634)
  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"; doi: 
            "10.1038/nature25442"; journalName: "Nature"; date: "2018-01-25- 
            25"; volume: "553"; issue: "7689"; pages: "496-500"
COMMENT     Sequence Label: CD40L-SrtA
FEATURES             Location/Qualifiers
     source          1..7634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1035..1044
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1041..1742
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1174..1192)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1442..1461
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1636..1659
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1752..1808
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3150..3173
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3377..3780
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4907..4923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4907..4923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4920..4942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4931..4947
                     /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(4955..4961,4962..4979,4980..4985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5000..5021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5138..5155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5309..5897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5389..5408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6068..6859,6860..6928))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6691..6710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6929..7033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7101..7119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7168..7190)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7314..7333
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7527..7549
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7541..7558
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7542..7558
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcaaggttag gaacagagag acaggagaat atgggccaaa caggatatct gtggtaagca
       61 gttcctgccc cggctcaggg ccaagaacag ttggaacagc agaatatggg ccaaacagga
      121 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
      181 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      241 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      301 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      361 ctccgattga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgca
      421 tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg actgcccacc
      481 tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc agggaccacc
      541 gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct ctgtctttgg
      601 gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttgg ctaactagat
      661 ctgtatctgg cggtcccgcg gaagaactga cgagttcgta ttcccggccg cagcccctgg
      721 gagacgtccc agcggcctcg ggggcccgtt ttgtggccca ttctgtatca gttaacctac
      781 ccgagtcgga ctttttggag ctccgccact gtccgagggg tacgtggctt tgttggggga
      841 cgagagacag agacacttcc cgcccccgtc tgaatttttg ctttcggttt tacgccgaaa
      901 ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
      961 tctgtatttg tctgaaaatt agctcgacaa agttaagtaa tagtccctct ctccaagctc
     1021 acttacaggc ggccgccacc atggtgagca agggcgagga ggtcatcaaa gagttcatgc
     1081 gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     1141 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc
     1201 ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg
     1261 tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt
     1321 gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc
     1381 tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg
     1441 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     1501 gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact
     1561 acctggtgga gttcaagacg atctacatgg ccaagaagcc cgtgcaactg cccggctact
     1621 actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac
     1681 agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac gagctgtaca
     1741 agggaagcgg agctactaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc
     1801 ctggaccgat agaaacatac agccaacctt cccccagatc cgtggcaact ggacttccag
     1861 cgagcatgaa gatttttatg tatttactta ctgttttcct tatcacccaa atgattggat
     1921 ctgtgctttt tgctgtgtat cttcatagaa gattggataa ggtcgaagag gaagtaaacc
     1981 ttcatgaaga ttttgtattc ataaaaaagc taaagagatg caacaaagga gaaggatctt
     2041 tatccttgct gaactgtgag gagatgagaa ggcaatttga agaccttgtc aaggatataa
     2101 cgttaaacaa agaagagaaa aaagaaaaca gctttgaaat gcaaagaggt gatgaggatc
     2161 ctcaaattgc agcacacgtt gtaagcgaag ccaacagtaa tgcagcatcc gttctacagt
     2221 gggccaagaa aggatattat accatgaaaa gcaacttggt aatgcttgaa aatgggaaac
     2281 agctgacggt taaaagagaa ggactctatt atgtctacac tcaagtcacc ttctgctcta
     2341 atcgggagcc ttcgagtcaa cgcccattca tcgtcggcct ctggctgaag cccagcagtg
     2401 gatctgagag aatcttactc aaggcggcaa atacccacag ttcctcccag ctttgcgagc
     2461 agcagtctgt tcacttgggc ggagtgtttg aattacaagc tggtgcttct gtgtttgtca
     2521 acgtgactga agcaagccaa gtgatccaca gagttggctt ctcatctttt ggcttactca
     2581 aactcaccgg tagcaccagc ggcagcggca agcccggcag cggcgagggc agcaccaagg
     2641 gcagtactag tggaagtggt aaacctggaa gtggagatgg gagtacgaaa ggaacgcgtc
     2701 aagctaaacc tcaaattccg aaagataaat cgaaagtggc aggctatatt gaaattccag
     2761 atgctgatat taaagaacca gtatatccag gaccagcaac atccgaacaa ttaaatagag
     2821 gtgtaagctt tgcagaagaa aatgaatcac tagatgatca aaatatttca attgcaggac
     2881 acactttcat tgaccgtccg aactatcaat ttacaaatct taaagcagcc aaaaaaggta
     2941 gtatggtgta ctttaaagtt ggtaatgaaa cacgtaagta taaaatgaca agtataagaa
     3001 acgttaagcc tacagatgta ggagttctag atgaacaaaa aggtaaagat aaacaattaa
     3061 cattaattac ttgtgatgat tacaatgaaa agacaggcgt ttgggaaacc cgtaaaatct
     3121 ttgtagctac agaagtcaaa ggatccggag actacaagga tgacgatgac aaataagtga
     3181 attcgagcat cttaccgcca tttattccca tatttgttct gtttttcttg atttgggtat
     3241 acatttaaat gttaataaaa caaaatggtg gggcaatcat ttacatttta tgggatatgt
     3301 aattactagt tcaggtgtat tgccacaaga caaacatgtt aagaaacttt cccgttattt
     3361 acgctctgtt cctgttaatc aacctctgga ttacaaaatt tgtgaaagat tgactgatat
     3421 tcttaactat gttgctcctt ttacgctgtg tggatatgct gctttaatgc ctctgtatca
     3481 tgctattgct tcccgtacgg ctttcgtttt ctcctccttg tataaatcct ggttgctgtc
     3541 tctttatgag gagttgtggc ccgttgtccg tcaacgtggc gtggtgtgct ctgtgtttgc
     3601 tgacgcaacc cccactggct ggggcattgc caccacctgt caactccttt ctgggacttt
     3661 cgctttcccc ctcccgatcg ccacggcaga actcatcgcc gcctgccttg cccgctgctg
     3721 gacaggggct aggttgctgg gcactgataa ttccgtggtg ttgtcgggga agctgacgtc
     3781 ctttccatgg ctgctcgcct gtgttgccaa ctggatcctg cgcgggacgt ccttctgcta
     3841 cgtcccttcg gctctcaatc cagcggacct cccttcccga ggccttctgc cggttctgcg
     3901 gcctctcccg cgtcttcgct ttcggcctcc gacgagtcgg atctcccttt gggccgcctc
     3961 cccgcctgtt tcgcctcggc gtccggtccg tgttgcttgg tcgtcacctg tgcagaattg
     4021 cgaaccatgg attccaccgt gaactttgtc tcctggcatg caaatcgtca acttggcatg
     4081 ccaagaatta attcggatcc aagcttaggc ctgctcgctt tcttgctgtc ccatttctat
     4141 taaaggttcc tttgttccct aagtccaact actaaactgg gggatattat gaagggcctt
     4201 gagcatctgg attctgccta gcgctaagct tcctaacacg agccatagat agaataaaag
     4261 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     4321 ctagcttaag taagccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt
     4381 tcagatcaag gttaggaaca gagagacagg agaatatggg ccaaacagga tatctgtggt
     4441 aagcagttcc tgccccggct cagggccaag aacagttgga acagcagaat atgggccaaa
     4501 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     4561 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     4621 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     4681 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     4741 cagtcctccg atagactgcg tcgcccgggg tacccgtatt cccaataaag cctcttgctg
     4801 tttgcatccg aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg
     4861 cccacctcgg gggtctttca ttctcgagag ctttggcgta atcatggtca tagctgtttc
     4921 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4981 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     5521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6361 tgcgcaacgt tgttgccatt gctgctggca tcgtggtgtc acgctcgtcg tttggtatgg
     6421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7021 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7081 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagctgcct
     7141 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     7201 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     7261 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagttact atgcggcatc
     7321 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     7381 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     7441 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     7501 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     7561 attagtactc tagcttaagt aagccatttt gcaaggcatg gaaaaataca taactgagaa
     7621 tagagaagtt caga
//
