LOCUS       sequence-320952-        6606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral (MSCV) expression of td Tomato fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6606)
  AUTHORS   Landry LG, Mann SE, Anderson AM, Nakayama M
  TITLE     Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation 
            Reporter-based Detection System.
  JOURNAL   Bio Protoc. 2021 Jan 20;11(2):e3883. doi: 10.21769/BioProtoc.3883. 
            eCollection 2021 Jan 20.
  PUBMED    33732772
REFERENCE   2  (bases 1 to 6606)
  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: "Bio 
            Protoc."; date: "2021-01-20"; volume: "11(2):e3883. doi"; pages: " 
            10.21769/BioProtoc.3883. eCollection 2021 Jan 20"
COMMENT     Sequence Label: pMSCVII-tdTomato
FEATURES             Location/Qualifiers
     source          1..6606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1420..1429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1426..2856
                     /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"
     LTR             3001..3515
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3677..3693
                     /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(3701..3707,3708..3725,3726..3731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3746..3767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3884..3901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4055..4643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4135..4154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4814..5605,5606..5674))
                     /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     5437..5456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5675..5779)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5847..5865
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5903..5925)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6025..6044
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6238..6260
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6388..6407
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tcagatctag ccaccatggt gagcaagggc
     1441 gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     1501 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1561 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     1621 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     1681 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     1741 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     1801 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1861 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     1921 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     1981 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     2041 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     2101 ctggggcatg gcaccggcag caccggcagc ggcagctccg gcaccgcctc ctccgaggac
     2161 aacaacatgg ccgtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg
     2221 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     2281 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     2341 ccccagttca tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac
     2401 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2461 ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg
     2521 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     2581 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     2641 caggccctga agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg
     2701 gccaagaagc ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc
     2761 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     2821 ctgttcctgt acggcatgga cgagctgtac aagtagctcg agcctgcagg catgcaagct
     2881 tcaggtagcc ggctaacgtt aacaaccggt acctctagaa ctatagctag catgcgcaaa
     2941 tttaaagcgc tgatatcgat aaaataaaag attttattta gtctccagaa aaagggggga
     3001 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     3061 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
     3121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
     3241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     3301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     3361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat
     3421 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     3481 gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag
     3541 aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt
     3601 ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct
     3661 ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3721 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3841 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4801 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4861 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4921 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4981 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5041 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5101 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5161 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5221 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5281 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5341 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5401 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5461 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5521 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5581 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5641 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5701 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5761 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5821 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     5881 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     5941 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     6001 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     6061 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     6121 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     6181 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     6241 agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa
     6301 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     6361 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     6421 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt
     6481 tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct
     6541 gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa
     6601 gggggg
//
