LOCUS       sequence-237108-        6930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMP71 vector expressing Tomato - P2A - flag - SrtA - PDGFR Tm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6930)
  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 6930)
  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: SrtA-PDGFR Tm
FEATURES             Location/Qualifiers
     source          1..6930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..57
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(95..117)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     217..236
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     430..452
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     444..461
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     445..461
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1572..1581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1578..2279
                     /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(1711..1729)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1979..1998
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2173..2196
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2289..2345
                     /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             2406..2429
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2901..3047
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    3251..3654
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             4160..4750
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4775..4791)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4775..4791)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4788..4810)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4799..4815
                     /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(4823..4829,4830..4847,4848..4853))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4868..4889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5006..5023)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5177..5765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5257..5276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5936..6727,6728..6796))
                     /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     6559..6578
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6797..6901)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
       61 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
      121 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
      181 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
      241 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      301 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      361 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      421 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattagta ctctagctta
      481 agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca
      541 aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
      601 tctgccccgg ctcagggcca agaacagttg gaacagcaga atatgggcca aacaggatat
      661 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
      721 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
      781 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
      841 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
      901 cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct gtttgtatcc
      961 gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact gcccacctcg
     1021 ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg gaccaccgac
     1081 ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg tctttgtgcg
     1141 tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttggcta actagatctg
     1201 tatctggcgg tcccgcggaa gaactgacga gttcgtattc ccggccgcag cccctgggag
     1261 acgtcccagc ggcctcgggg gcccgttttg tggcccattc tgtatcagtt aacctacccg
     1321 agtcggactt tttggagctc cgccactgtc cgaggggtac gtggctttgt tgggggacga
     1381 gagacagaga cacttcccgc ccccgtctga atttttgctt tcggttttac gccgaaaccg
     1441 cgccgcgcgt cttgtctgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct
     1501 gtatttgtct gaaaattagc tcgacaaagt taagtaatag tccctctctc caagctcact
     1561 tacaggcggc cgccaccatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct
     1621 tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     1681 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
     1741 tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga
     1801 agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg
     1861 agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc
     1921 aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc
     1981 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg
     2041 acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc
     2101 tggtggagtt caagacgatc tacatggcca agaagcccgt gcaactgccc ggctactact
     2161 acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2221 acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag ctgtacaagg
     2281 gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg
     2341 gaccggtacc gtgcacgctg ctcctgctgt tggcggccgc cctggctccg actcagaccc
     2401 gcgccgacta caaggatgac gatgacaaag gttcaggtct cgagcaagct aaacctcaaa
     2461 ttccgaaaga taaatcgaaa gtggcaggct atattgaaat tccagatgct gatattaaag
     2521 aaccagtata tccaggacca gcaacatccg aacaattaaa tagaggtgta agctttgcag
     2581 aagaaaatga atcactagat gatcaaaata tttcaattgc aggacacact ttcattgacc
     2641 gtccgaacta tcaatttaca aatcttaaag cagccaaaaa aggtagtatg gtgtacttta
     2701 aagttggtaa tgaaacacgt aagtataaaa tgacaagtat aagaaacgtt aagcctacag
     2761 atgtaggagt tctagatgaa caaaaaggta aagataaaca attaacatta attacttgtg
     2821 atgattacaa tgaaaagaca ggcgtttggg aaacccgtaa aatctttgta gctacagaag
     2881 tcaaaggtgg tggcggttca gctgtgggcc aggacacgca ggaggtcatc gtggtgccac
     2941 actccttgcc ctttaaggtg gtggtgatct cagccatcct ggccctggtg gtgctcacca
     3001 tcatctccct tatcatcctc atcatgcttt ggcagaagaa gccacgttaa gtgaattcga
     3061 gcatcttacc gccatttatt cccatatttg ttctgttttt cttgatttgg gtatacattt
     3121 aaatgttaat aaaacaaaat ggtggggcaa tcatttacat tttatgggat atgtaattac
     3181 tagttcaggt gtattgccac aagacaaaca tgttaagaaa ctttcccgtt atttacgctc
     3241 tgttcctgtt aatcaacctc tggattacaa aatttgtgaa agattgactg atattcttaa
     3301 ctatgttgct ccttttacgc tgtgtggata tgctgcttta atgcctctgt atcatgctat
     3361 tgcttcccgt acggctttcg ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3421 tgaggagttg tggcccgttg tccgtcaacg tggcgtggtg tgctctgtgt ttgctgacgc
     3481 aacccccact ggctggggca ttgccaccac ctgtcaactc ctttctggga ctttcgcttt
     3541 ccccctcccg atcgccacgg cagaactcat cgccgcctgc cttgcccgct gctggacagg
     3601 ggctaggttg ctgggcactg ataattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3661 atggctgctc gcctgtgttg ccaactggat cctgcgcggg acgtccttct gctacgtccc
     3721 ttcggctctc aatccagcgg acctcccttc ccgaggcctt ctgccggttc tgcggcctct
     3781 cccgcgtctt cgctttcggc ctccgacgag tcggatctcc ctttgggccg cctccccgcc
     3841 tgtttcgcct cggcgtccgg tccgtgttgc ttggtcgtca cctgtgcaga attgcgaacc
     3901 atggattcca ccgtgaactt tgtctcctgg catgcaaatc gtcaacttgg catgccaaga
     3961 ataattcgga tccaagctta ggcctgctcg ctttcttgct gtcccatttc tattaaaggt
     4021 tcctttgttc cctaagtcca actactaaac tgggggatat tatgaagggc cttgagcatc
     4081 tggattctgc ctagcgctaa gcttaacacg agccatagat agaataaaag attttattta
     4141 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     4201 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     4261 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     4321 tgccccggct cagggccaag aacagttgga acagcagaat atgggccaaa caggatatct
     4381 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     4441 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     4501 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     4561 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     4621 atagactgcg tcgcccgggt acccgtgttc tcaataaacc ctcttgcagt tgcatccgac
     4681 tcgtggtctc gctgttcctt gggagggtct cctctgagtg attgactgcc cacctcgggg
     4741 gtctttcatt ctcgagcagc ttggcgtaat aatggtcata gctgtttcct gtgtgaaatt
     4801 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4861 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4921 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4981 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5041 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5101 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5161 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5221 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5281 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5341 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5401 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5461 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5521 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5581 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5641 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5701 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5761 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5821 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5881 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5941 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6001 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6061 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6121 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6181 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6241 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6301 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6361 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6421 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6481 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6541 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6601 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6661 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6721 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6781 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6841 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6901 gcacatttcc ccgaaaagtg ccacctgacg
//
