LOCUS       sequence-208621-        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid containing tdTomato for labeling of cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7871)
  AUTHORS   Kleeman B, Olsson A, Newkold T, Kofron M, DeLay M, Hildeman D, 
            Grimes HL
  TITLE     A guide to choosing fluorescent protein combinations for flow 
            cytometric analysis based on spectral overlap.
  JOURNAL   Cytometry A. 2018 Mar 13. doi: 10.1002/cyto.a.23360.
  PUBMED    29533508
REFERENCE   2  (bases 1 to 7871)
  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"; journalName: "Cytometry 
            A. 2018 Mar 13. doi: 10.1002/cyto.a.23360."
COMMENT     Sequence Label: pMSCVpuro-tdTomato
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1410..1466
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1474..1483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1480..2910
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     promoter        3045..3544
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3080..3103)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3477..3496
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3565..4164
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3565..3584)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4061..4081
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4264..4778
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4940..4956
                     /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(4964..4970,4971..4988,4989..4994))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5009..5030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5147..5164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5318..5906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5398..5417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6077..6868,6869..6937))
                     /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     6700..6719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6938..7042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7110..7128
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7166..7188)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7288..7307
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7501..7523
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7651..7670
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7870..7871,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctcgagc tcaagcttcg aattctgcag
     1441 tcgacggtac cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     1501 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
     1561 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1621 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
     1681 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
     1741 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
     1801 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
     1861 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1921 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag
     1981 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc
     2041 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag
     2101 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgggg
     2161 catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga ggacaacaac
     2221 atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     2281 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2341 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     2401 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     2461 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     2521 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     2581 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2641 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     2701 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     2761 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     2821 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     2881 ctgtacggca tggacgagct gtacaagtaa gcggccgcga ctctagatca taatcagcca
     2941 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3001 gaaacataaa atgaatgcaa ttgttgttgt taacgaattc taccgggtag gggaggcgct
     3061 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     3121 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     3181 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     3241 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     3301 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     3361 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     3421 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
     3481 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     3541 ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc
     3601 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     3661 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     3721 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     3781 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     3841 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     3901 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     3961 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4021 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4081 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4141 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg
     4201 agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca gaaaaagggg
     4261 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     4321 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     4381 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4441 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     4501 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     4561 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     4621 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc
     4681 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     4741 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg
     4801 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact
     4861 gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga
     4921 gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4981 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5041 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5101 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5161 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5221 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5281 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5341 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5401 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5461 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5521 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5581 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5641 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5701 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5761 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5821 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5881 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5941 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6001 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6061 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6121 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6181 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6241 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6301 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6361 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6421 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6481 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6541 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6601 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6661 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6721 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6781 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6841 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6901 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6961 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7021 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7081 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     7141 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     7201 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     7261 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     7321 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     7381 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7441 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     7501 cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg agatggcgcc
     7561 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     7621 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     7681 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc gattagtcca
     7741 atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac aatatcacca
     7801 gctgaagcct atagagtacg agccatagat aaaataaaag attttattta gtctccagaa
     7861 aaagggggga a
//
