LOCUS       sequence-237104-        7209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMP71 vector expressing GFP - P2A  - G5 - myc - CD40.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7209)
  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 7209)
  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: G5-CD40
FEATURES             Location/Qualifiers
     source          1..7209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..93
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     287..309
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     301..318
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     302..318
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1495..1497,1498..2208)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1537..1558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1798..1817)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2145..2166
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2218..2274
                     /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             2356..2385
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3387..3790
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             4296..4886
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4911..4927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4911..4927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4924..4946)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4935..4951
                     /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(4959..4965,4966..4983,4984..4989))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5004..5025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5142..5159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5313..5901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5393..5412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6072..6863,6864..6932))
                     /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     6695..6714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6933..7037)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7105..7123
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7161..7183)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
       61 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
      121 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
      181 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      241 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      301 tgtaaaacga cggccagtga attagtactc tagcttaagt aacgccattt tgcaaggcat
      361 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      421 gaatatgggc caaacaggat atctgtggta agcagtttct gccccggctc agggccaaga
      481 acagttggaa cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
      541 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
      601 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
      661 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
      721 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
      781 cccgtattcc caataaagcc tcttgctgtt tgtatccgaa tcgtggactc gctgatcctt
      841 gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc
      901 accgagattt ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg
      961 ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt
     1021 tgcgcctgcg tctgtactag ttggctaact agatctgtat ctggcggtcc cgcggaagaa
     1081 ctgacgagtt cgtattcccg gccgcagccc ctgggagacg tcccagcggc ctcgggggcc
     1141 cgttttgtgg cccattctgt atcagttaac ctacccgagt cggacttttt ggagctccgc
     1201 cactgtccga ggggtacgtg gctttgttgg gggacgagag acagagacac ttcccgcccc
     1261 cgtctgaatt tttgctttcg gttttacgcc gaaaccgcgc cgcgcgtctt gtctgctgca
     1321 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagctcg
     1381 acaaagttaa gtaatagtcc ctctctccaa gctcacttac aggcggccgc caccatgctg
     1441 tgctgtatga gaagaaccaa acaggttgaa aagaatgatg aggaccaaaa gatcgtgagc
     1501 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1561 aacggccaca agttcagcgt gtctggcgag ggcgagggcg atgccaccta cggcaagctg
     1621 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1681 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1741 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1801 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1861 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1921 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1981 gcgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     2041 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2101 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2161 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg aagcggagct
     2221 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg accggtgtct
     2281 ttgcctcggc tgtgcgcgct atggggctgc ttgttgacag cggtccatct agggcagtgt
     2341 ggaggcggag gaggcgaaca aaaacttatt tctgaagaag atctgacgcg ttgcagtgac
     2401 aaacagtacc tccacgatgg ccagtgctgt gatttgtgcc agccaggaag ccgactgaca
     2461 agccactgca cagctcttga gaagacccaa tgccacccat gtgactcagg cgaattctca
     2521 gcccagtgga acagggagat tcgctgtcac cagcacagac actgtgaacc caatcaaggg
     2581 cttcgggtta agaaggaggg caccgcagaa tcagacactg tctgtacctg taaggaagga
     2641 caacactgca ccagcaagga ttgcgaggca tgtgctcagc acacgccctg tatccctggc
     2701 tttggagtta tggagatggc cactgagacc actgataccg tctgtcatcc ctgcccagtc
     2761 ggcttcttct ccaatcagtc atcacttttc gaaaagtgtt atccctggac aagctgtgag
     2821 gataagaact tggaggtcct acagaaagga acgagtcaga ctaatgtcat ctgtggttta
     2881 aagtcccgga tgcgagccct gctggtcatt cctgtcgtga tgggcatcct catcaccatt
     2941 ttcggggtgt ttctctatat caaaaaggtg gtcaagaaac caaaggataa tgagatctta
     3001 ccccctgcgg ctcgacggca agatccccag gagatggaag attatcccgg tcataacacc
     3061 gctgctccag tgcaggagac gctgcacggg tgtcagcctg tcacacagga ggatggtaaa
     3121 gagagtcgca tctcagtgca ggagcggcag gtgacagaca gcatagcctt gaggcccctg
     3181 gtctaagtga attcgagcat cttaccgcca tttattccca tatttgttct gtttttcttg
     3241 atttgggtat acatttaaat gttaataaaa caaaatggtg gggcaatcat ttacatttta
     3301 tgggatatgt aattactagt tcaggtgtat tgccacaaga caaacatgtt aagaaacttt
     3361 cccgttattt acgctctgtt cctgttaatc aacctctgga ttacaaaatt tgtgaaagat
     3421 tgactgatat tcttaactat gttgctcctt ttacgctgtg tggatatgct gctttaatgc
     3481 ctctgtatca tgctattgct tcccgtacgg ctttcgtttt ctcctccttg tataaatcct
     3541 ggttgctgtc tctttatgag gagttgtggc ccgttgtccg tcaacgtggc gtggtgtgct
     3601 ctgtgtttgc tgacgcaacc cccactggct ggggcattgc caccacctgt caactccttt
     3661 ctgggacttt cgctttcccc ctcccgatcg ccacggcaga actcatcgcc gcctgccttg
     3721 cccgctgctg gacaggggct aggttgctgg gcactgataa ttccgtggtg ttgtcgggga
     3781 agctgacgtc ctttccatgg ctgctcgcct gtgttgccaa ctggatcctg cgcgggacgt
     3841 ccttctgcta cgtcccttcg gctctcaatc cagcggacct cccttcccga ggccttctgc
     3901 cggttctgcg gcctctcccg cgtcttcgct ttcggcctcc gacgagtcgg atctcccttt
     3961 gggccgcctc cccgcctgtt tcgcctcggc gtccggtccg tgttgcttgg tcgtcacctg
     4021 tgcagaattg cgaaccatgg attccaccgt gaactttgtc tcctggcatg caaatcgtca
     4081 acttggcatg ccaagaataa ttcggatcca agcttaggcc tgctcgcttt cttgctgtcc
     4141 catttctatt aaaggttcct ttgttcccta agtccaacta ctaaactggg ggatattatg
     4201 aagggccttg agcatctgga ttctgcctag cgctaagctt aacacgagcc atagatagaa
     4261 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     4321 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
     4381 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
     4441 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gttggaacag cagaatatgg
     4501 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4561 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     4621 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     4681 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     4741 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgttctcaa taaaccctct
     4801 tgcagttgca tccgactcgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     4861 actgcccacc tcgggggtct ttcattctcg agcagcttgg cgtaatcatg gtcatagctg
     4921 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4981 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5041 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5101 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5161 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5221 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5281 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5341 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5401 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5461 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5521 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5581 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5641 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5701 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5761 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5821 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5881 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5941 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6001 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6061 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6181 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6361 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6661 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6721 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6841 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6901 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6961 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7021 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     7081 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     7141 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     7201 ctgtaagcg
//
