LOCUS       sequence-322031-        7675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector to overexpress the murine CD3 (TCR) components CD3
            gamma, CD delta, CD epsilon and TCR zeta and surface marker CD90.2, 
            separated by T2; F2A; E2A; P2A respectively, under control of the 
            hFTH1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7675)
  AUTHORS   van der Donk LEH, van der Spek J, van Duivenvoorde T, Ten Brink MS, 
            Geijtenbeek TBH, Kuijl CP, van Heijst JWJ, Ates LS
  TITLE     An optimized retroviral toolbox for overexpression and genetic 
            perturbation of primary lymphocytes.
  JOURNAL   Biol Open. 2022 Feb 15;11(2):bio059032. doi: 10.1242/bio.059032. 
            Epub 2022 Mar 1.
  PUBMED    35229875
REFERENCE   2  (bases 1 to 7675)
  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: "Biol 
            Open."; date: "2022-02-15"; volume: "11(2):bio059032. doi"; pages: "
            10.1242/bio.059032. Epub 2022 Mar 1"
COMMENT     Sequence Label: pMX-hFTH1-CD3-P2A-CD90.2
FEATURES             Location/Qualifiers
     source          1..7675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1898
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1836..1852
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      2311..2320
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2878..2931
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(5779..5796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5950..6538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6030..6049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6702..7493,7494..7562))
                     /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     7325..7344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7563..7667)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggccggtg gtacctcacc cttaccgagt cggcgacaca gtgtgggtcc
     1741 gccgacacca gactaagaac ctagaacctc gctggaaagg accttacaca gtcctgctga
     1801 ccacccccac cgccctcaaa gtagacggca tcgcagcttg gatacacgcc gcccacgtga
     1861 aggctgccga ccccgggggt ggaccatcct ctagactgcc ggatctagct agttaattaa
     1921 tccgccagag cgcgcgaggg cctccaccgg ccgcccctcc cccacagcag gggcggggtc
     1981 ccgcgcccac cggaaggagc gggctcgggg cgggcggcgc tgattggccg gggcgggcct
     2041 gacgccgacg cggctataag agaccacaag cgacccgcag ggccagacgt tcttcgccga
     2101 gagtcgtcgg ggtttcctgc ttcaacagtg cttggacgga acccggcgct cgttccccac
     2161 cccggccggc cgcccatagc cagccctccg tcacctcttc accgcaccct cggactgccc
     2221 caaggccccc gccgccgctc cagcgccgcg cagccaccgc cgccgccgcc gcctctcctt
     2281 agtcgccgcc ggatccggtc gtgcggccgc gccaccatgg aacagagaaa aggcctggcc
     2341 ggcctgttcc tggttatcag tctgctgcag ggcacagtgg cccagaccaa caaggctaag
     2401 aacctggtgc aggtggacgg ctctagaggc gacggatctg tgctgctgac atgtggcctg
     2461 accgacaaga ccatcaagtg gctgaaggac ggctccatca tcagccctct gaacgccacc
     2521 aagaacacct ggaacctggg caacaacgcc aaggacccca gaggcaccta tcagtgccag
     2581 ggcgccaaag agacaagcaa ccctctgcag gtctactaca gaatgtgcga gaactgcatc
     2641 gagctgaaca tcggcaccat cagcggcttc atcttcgccg aagtgatcag catcttcttt
     2701 ctggccctgg gcgtgtacct gatcgctgga caagatggcg tgcggcagag cagagccagc
     2761 gataagcaga cactgctgca gaacgagcag ctgtaccagc ctctgaagga cagagagtac
     2821 gaccagtaca gccacctcca gggcaaccag ctgcggaaga agggatctgg ccaattggaa
     2881 ggcagaggct ctcttcttac atgcggcgac gtcgaggaaa acccaggacc tatggaacac
     2941 tctggcatcc tggctagcct gatcctgatt gccgttctgc ctcaaggcag ccccttcaag
     3001 atccaagtga ccgagtacga ggacaaggtg ttcgtgacct gcaacaccag cgtgatgcac
     3061 ctggatggca ccgtggaagg atggttcgcc aagaacaaga ccctgaacct cggcaagggc
     3121 gtgctggacc ctagaggcat ctacctgtgt aacggcacag agcagctggc caaggtggtg
     3181 tctagtgtgc aggtccacta tcggatgtgt cagaactgcg tggaactgga cagcggcaca
     3241 atggccggcg tgatcttcat cgacctgatc gctaccctgc tgctggcact gggagtgtat
     3301 tgcttcgctg gccacgagac aggcagacct agcggagctg ctgaagttca ggccctgctg
     3361 aagaatgaac agctctatca gcccctgcgc gacagagagg atacccagta ctctagactc
     3421 ggcggcaact ggcccagaaa caagaaatct ggaagcggca cgcgtgtcag acagaccctg
     3481 aacttcgatc tgcttagact ggccggggac gtcgagtcta atccaggacc aatgcggtgg
     3541 aacaccttct ggggcatcct gtgtctgtct ctgctggctg tgggcacctg tcaggatgac
     3601 gctgagaaca tcgagtataa ggtgtccatc tccggcacca gcgtcgagct gacttgtcct
     3661 ctggactccg acgagaacct gaagtgggag aagaacggcc aagagctgcc tcagaagcac
     3721 gacaagcacc tggtgctgca ggacttcagc gaggtggaag atagcggcta ctacgtgtgc
     3781 tacacccctg ccagcaacaa gaacacatac ctgtacctga aggctcgcgt gtgcgagtac
     3841 tgtgtcgagg tggacctgac agccgtggct atcatcatca tcgtggacat ctgcatcacc
     3901 ctgggcctgc tgatggtcat ctactactgg tccaagaacc ggaaggccaa ggccaagcct
     3961 gtgacaagag gaaccggcgc tggaagcaga ccaagaggcc agaacaaaga aagacctcct
     4021 cctgtgccta atcctgacta cgagcccatc cggaagggcc agagagatct gtactctggc
     4081 ctgaaccaga gggccgtggg ttctggcgca tgccagtgta ccaactatgc tctcctgaga
     4141 ctcgcaggcg acgttgagag taatccaggg cctatgaagt ggaaagtgtc tgtgctggcc
     4201 tgcatcctgc atgttcgatt ccctggcgct gaggcccagt cttttggact gctggacccc
     4261 aagctgtgct acctgctgga cggcattctg tttatttatg gcgtgatcat caccgctctg
     4321 tacctgcggg ccaagttcag cagaagcgct gagacagctg ccaatctgca ggaccctaac
     4381 cagctgtaca acgagctgaa tctggggcgc agagaagagt acgatgtgct ggaaaagaag
     4441 agagccagag atcccgagat gggcggcaaa cagcagagaa ggcggaatcc tcaagaaggc
     4501 gtgtacaacg ccctgcagaa agataagatg gccgaggcct acagcgagat cggcacaaag
     4561 ggcgaacgca gaagaggcaa gggacacgat ggactgtacc agggcctgtc cacagccaca
     4621 aaggacacat acgatgccct gcacatgcag acactggccc ctagaggcag cggcgtcgac
     4681 gccacaaact tcagcctgct gagacaggct ggcgacgtgg aagagaatcc tggacctatg
     4741 aaccccgcca tctctgtggc cctgctgctg tctgtgctgc aggtgtccag aggccagaaa
     4801 gtgaccagcc tgaccgcctg cctcgtgaac cagaacctga gactggactg cagacacgag
     4861 aacaacacca aggacaacag catccagcac gagttctccc tgacccgcga gaagagaaaa
     4921 cacgtgctga gcggcaccct gggcatcccc gagcacacct acagatccag agtgaccctg
     4981 agcaaccagc cctacatcaa ggtgctgacc ctggccaact tcaccacaaa ggacgagggc
     5041 gactacttct gcgaactgca ggtgtcaggc gccaacccca tgagcagcaa caagagcatc
     5101 agcgtgtaca gagacaagct cgtgaagtgc ggcggcatca gcctgctggt gcagaacacc
     5161 tcttggatgc tgttgctgct gctgagcctg agcctgctgc aggccctgga cttcatctct
     5221 ctgtagcctg caggtttctg aagcttgagt acatcgataa aataaaagat tttatttagt
     5281 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agctagagaa
     5341 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     5401 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     5461 gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc
     5521 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     5581 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc
     5641 ttgaagtgtc gagcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat
     5701 taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5761 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5821 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5881 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5941 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6001 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6061 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6121 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6181 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6241 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6301 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6361 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6421 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6481 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6541 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6601 gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc ggccggccgc
     6661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6721 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6781 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6841 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6901 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6961 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7021 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7081 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7141 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7201 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7261 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7321 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7381 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7441 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7501 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7561 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7621 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttc
//
