LOCUS       sequence-472361-        6787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMP71 backbone. Expresses Thy1.1 with N-terminal 5xGly (G5).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6787)
  AUTHORS   Nakandakari-Higa S, Walker S, Canesso MCC, van der Heide V, 
            Chudnovskiy A, Kim DY, Jacobsen JT, Parsa R, Bilanovic J, Parigi SM,
            Fiedorczuk K, Fuchs E, Bilate AM, Pasqual G, Mucida D, Kamphorst AO,
            Pritykin Y, Victora GD
  TITLE     Universal recording of immune cell interactions in vivo.
  JOURNAL   Nature. 2024 Mar;627(8003):399-406. doi: 10.1038/s41586-024-07134-4.
            Epub 2024 Mar 6.
  PUBMED    38448581
REFERENCE   2  (bases 1 to 6787)
  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/s41586-024-07134-4"; journalName: "Nature"; date: 
            "2024-03"; volume: "627"; issue: "8003"; pages: "399-406"
COMMENT     Sequence Label: Thy1.1-G5
FEATURES             Location/Qualifiers
     source          1..6787
                     /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             join(1041..1043,1044..1046,1047..1757)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1086..1107)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1347..1366)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1694..1715
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1767..1823
                     /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             1905..1934
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2571..2974
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             3480..4070
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4095..4111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4095..4111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4108..4130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4119..4135
                     /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(4143..4149,4150..4167,4168..4173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4188..4209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4326..4343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4497..5085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4577..4596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5256..6047,6048..6116))
                     /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     5879..5898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6117..6221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6289..6307
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6345..6367)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6467..6486
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6680..6702
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6694..6711
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6695..6711
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
       61 gtttctgccc 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 ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgta
      421 tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg actgcccacc
      481 tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc agggaccacc
      541 gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct ctgtctttgt
      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 gctgttcacc ggggtggtgc
     1081 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tctggcgagg
     1141 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1201 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1261 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1321 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1381 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1441 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1501 tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacatcgagg
     1561 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1621 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     1681 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1741 tggacgagct gtacaaggga agcggagcta ctaacttcag cctgctgaag caggctggag
     1801 acgtggagga gaaccctgga ccggtgtctt tgcctcggct gtgcgcgcta tggggctgct
     1861 tgttgacagc ggtccatcta gggcagtgtg gaggcggagg aggcgaacaa aaacttattt
     1921 ctgaagaaga tctgacgcgt cagaaggtga ccagcctgac agcctgcctg gtgaaccaaa
     1981 accttcgcct ggactgccgc catgagaata acaccaagga taactccatc cagcatgagt
     2041 tcagcctgac ccgagagaag aggaagcacg tgctctcagg caccctcggg atacccgagc
     2101 acacgtaccg ctcccgcgtc accctctcca accagcccta tatcaaggtc cttaccctag
     2161 ccaacttcac caccaaggat gagggcgact acttttgtga gcttcgagtc tcgggcgcga
     2221 atcccatgag ctccaataaa agtatcagtg tgtatagaga caaactggtc aagtgtggcg
     2281 gcataagcct gctggttcag aacacatcct ggatgctgct gctgctgctt tccctctccc
     2341 tcctccaagc cctggacttc atttctctgt gagaattcga gcatcttacc gccatttatt
     2401 cccatatttg ttctgttttt cttgatttgg gtatacattt aaatgttaat aaaacaaaat
     2461 ggtggggcaa tcatttacat tttatgggat atgtaattac tagttcaggt gtattgccac
     2521 aagacaaaca tgttaagaaa ctttcccgtt atttacgctc tgttcctgtt aatcaacctc
     2581 tggattacaa aatttgtgaa agattgactg atattcttaa ctatgttgct ccttttacgc
     2641 tgtgtggata tgctgcttta atgcctctgt atcatgctat tgcttcccgt acggctttcg
     2701 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2761 tccgtcaacg tggcgtggtg tgctctgtgt ttgctgacgc aacccccact ggctggggca
     2821 ttgccaccac ctgtcaactc ctttctggga ctttcgcttt ccccctcccg atcgccacgg
     2881 cagaactcat cgccgcctgc cttgcccgct gctggacagg ggctaggttg ctgggcactg
     2941 ataattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     3001 ccaactggat cctgcgcggg acgtccttct gctacgtccc ttcggctctc aatccagcgg
     3061 acctcccttc ccgaggcctt ctgccggttc tgcggcctct cccgcgtctt cgctttcggc
     3121 ctccgacgag tcggatctcc ctttgggccg cctccccgcc tgtttcgcct cggcgtccgg
     3181 tccgtgttgc ttggtcgtca cctgtgcaga attgcgaacc atggattcca ccgtgaactt
     3241 tgtctcctgg catgcaaatc gtcaacttgg catgccaaga ataattcgga tccaagctta
     3301 ggcctgctcg ctttcttgct gtcccatttc tattaaaggt tcctttgttc cctaagtcca
     3361 actactaaac tgggggatat tatgaagggc cttgagcatc tggattctgc ctagcgctaa
     3421 gcttaacacg agccatagat agaataaaag attttattta gtctccagaa aaagggggga
     3481 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     3541 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
     3601 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3661 aacagttgga acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3721 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga
     3781 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     3841 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     3901 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     3961 acccgtgttc tcaataaacc ctcttgcagt tgcatccgac tcgtggtctc gctgttcctt
     4021 gggagggtct cctctgagtg attgactgcc cacctcgggg gtctttcatt ctcgagcagc
     4081 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4141 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4201 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4261 ctgcattaat gaatctgcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4321 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4381 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4441 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4501 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4561 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4621 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4681 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4741 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4801 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4861 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4921 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4981 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5041 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5101 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5161 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5221 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5281 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5341 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5401 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5461 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5521 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5581 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5641 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5701 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5761 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5821 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5881 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5941 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6001 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6061 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6121 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6181 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6241 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6301 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6361 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6421 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6481 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6541 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6601 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6661 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattagta
     6721 ctctagctta agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa
     6781 gttcaga
//
