LOCUS       sequence-237106-        7593 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 7593)
  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 7593)
  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..7593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..24
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     218..240
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     232..249
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     233..249
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1360..1369
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1366..2067
                     /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(1499..1517)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1767..1786
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1961..1984
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2077..2133
                     /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             3475..3498
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3702..4105
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             4611..5201
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5226..5242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5226..5242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5239..5261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5250..5266
                     /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(5274..5280,5281..5298,5299..5304))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5319..5340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5457..5474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5628..6216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5708..5727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6387..7178,7179..7247))
                     /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     7010..7029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7248..7352)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7420..7438
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7476..7498)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
       61 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
      121 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      181 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      241 acggccagtg aattagtact ctagcttaag taacgccatt ttgcaaggca tggaaaatac
      301 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
      361 ccaaacagga tatctgtggt aagcagtttc tgccccggct cagggccaag aacagttgga
      421 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
      481 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
      541 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
      601 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
      661 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
      721 ccaataaagc ctcttgctgt ttgtatccga atcgtggact cgctgatcct tgggagggtc
      781 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
      841 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
      901 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
      961 gtctgtacta gttggctaac tagatctgta tctggcggtc ccgcggaaga actgacgagt
     1021 tcgtattccc ggccgcagcc cctgggagac gtcccagcgg cctcgggggc ccgttttgtg
     1081 gcccattctg tatcagttaa cctacccgag tcggactttt tggagctccg ccactgtccg
     1141 aggggtacgt ggctttgttg ggggacgaga gacagagaca cttcccgccc ccgtctgaat
     1201 ttttgctttc ggttttacgc cgaaaccgcg ccgcgcgtct tgtctgctgc agcatcgttc
     1261 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagctc gacaaagtta
     1321 agtaatagtc cctctctcca agctcactta caggcggccg ccaccatggt gagcaagggc
     1381 gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     1441 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1501 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     1561 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     1621 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     1681 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     1741 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1801 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     1861 ctgaagctga aggacggcgg ccactacctg gtggagttca agacgatcta catggccaag
     1921 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     1981 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     2041 ctgtacggca tggacgagct gtacaaggga agcggagcta ctaacttcag cctgctgaag
     2101 caggctggag acgtggagga gaaccctgga ccgatagaaa catacagcca accttccccc
     2161 agatccgtgg caactggact tccagcgagc atgaagattt ttatgtattt acttactgtt
     2221 ttccttatca cccaaatgat tggatctgtg ctttttgctg tgtatcttca tagaagattg
     2281 gataaggtcg aagaggaagt aaaccttcat gaagattttg tattcataaa aaagctaaag
     2341 agatgcaaca aaggagaagg atctttatcc ttgctgaact gtgaggagat gagaaggcaa
     2401 tttgaagacc ttgtcaagga tataacgtta aacaaagaag agaaaaaaga aaacagcttt
     2461 gaaatgcaaa gaggtgatga ggatcctcaa attgcagcac acgttgtaag cgaagccaac
     2521 agtaatgcag catccgttct acagtgggcc aagaaaggat attataccat gaaaagcaac
     2581 ttggtaatgc ttgaaaatgg gaaacagctg acggttaaaa gagaaggact ctattatgtc
     2641 tacactcaag tcaccttctg ctctaatcgg gagccttcga gtcaacgccc attcatcgtc
     2701 ggcctctggc tgaagcccag cagtggatct gagagaatct tactcaaggc ggcaaatacc
     2761 cacagttcct cccagctttg cgagcagcag tctgttcact tgggcggagt gtttgaatta
     2821 caagctggtg cttctgtgtt tgtcaacgtg actgaagcaa gccaagtgat ccacagagtt
     2881 ggcttctcat cttttggctt actcaaactc accggtagca ccagcggcag cggcaagccc
     2941 ggcagcggcg agggcagcac caagggcagt actagtggaa gtggtaaacc tggaagtgga
     3001 gatgggagta cgaaaggaac gcgtcaagct aaacctcaaa ttccgaaaga taaatcgaaa
     3061 gtggcaggct atattgaaat tccagatgct gatattaaag aaccagtata tccaggacca
     3121 gcaacatccg aacaattaaa tagaggtgta agctttgcag aagaaaatga atcactagat
     3181 gatcaaaata tttcaattgc aggacacact ttcattgacc gtccgaacta tcaatttaca
     3241 aatcttaaag cagccaaaaa aggtagtatg gtgtacttta aagttggtaa tgaaacacgt
     3301 aagtataaaa tgacaagtat aagaaacgtt aagcctacag atgtaggagt tctagatgaa
     3361 caaaaaggta aagataaaca attaacatta attacttgtg atgattacaa tgaaaagaca
     3421 ggcgtttggg aaacccgtaa aatctttgta gctacagaag tcaaaggatc cggagactac
     3481 aaggatgacg atgacaaata agtgaattcg agcatcttac cgccatttat tcccatattt
     3541 gttctgtttt tcttgatttg ggtatacatt taaatgttaa taaaacaaaa tggtggggca
     3601 atcatttaca ttttatggga tatgtaatta ctagttcagg tgtattgcca caagacaaac
     3661 atgttaagaa actttcccgt tatttacgct ctgttcctgt taatcaacct ctggattaca
     3721 aaatttgtga aagattgact gatattctta actatgttgc tccttttacg ctgtgtggat
     3781 atgctgcttt aatgcctctg tatcatgcta ttgcttcccg tacggctttc gttttctcct
     3841 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtccgtcaac
     3901 gtggcgtggt gtgctctgtg tttgctgacg caacccccac tggctggggc attgccacca
     3961 cctgtcaact cctttctggg actttcgctt tccccctccc gatcgccacg gcagaactca
     4021 tcgccgcctg ccttgcccgc tgctggacag gggctaggtt gctgggcact gataattccg
     4081 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccaactgga
     4141 tcctgcgcgg gacgtccttc tgctacgtcc cttcggctct caatccagcg gacctccctt
     4201 cccgaggcct tctgccggtt ctgcggcctc tcccgcgtct tcgctttcgg cctccgacga
     4261 gtcggatctc cctttgggcc gcctccccgc ctgtttcgcc tcggcgtccg gtccgtgttg
     4321 cttggtcgtc acctgtgcag aattgcgaac catggattcc accgtgaact ttgtctcctg
     4381 gcatgcaaat cgtcaacttg gcatgccaag aataattcgg atccaagctt aggcctgctc
     4441 gctttcttgc tgtcccattt ctattaaagg ttcctttgtt ccctaagtcc aactactaaa
     4501 ctgggggata ttatgaaggg ccttgagcat ctggattctg cctagcgcta agcttaacac
     4561 gagccataga tagaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     4621 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
     4681 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
     4741 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagttgg
     4801 aacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4861 gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca
     4921 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     4981 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     5041 acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgtt
     5101 ctcaataaac cctcttgcag ttgcatccga ctcgtggtct cgctgttcct tgggagggtc
     5161 tcctctgagt gattgactgc ccacctcggg ggtctttcat tctcgagcag cttggcgtaa
     5221 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5281 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5341 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5401 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5461 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5521 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5581 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5641 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5701 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5761 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5821 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5881 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5941 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6001 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6061 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6121 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6181 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6241 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6301 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6361 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6421 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6481 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6541 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6601 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6661 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6721 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6781 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6841 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6901 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6961 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7021 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7081 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7141 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7201 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7261 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7321 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7381 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7441 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7501 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     7561 gcgggtgttg gcgggtgtcg gggctggctt aac
//
