LOCUS       sequence-322011-        5644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for knockdown of murine CD247A (TCR zeta) and 
            expression of a CD90.2 surface marker under control of the LTR 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5644)
  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 5644)
  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: pMY-CD90.2-Cd247A_miR
FEATURES             Location/Qualifiers
     source          1..5644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             1103..1519
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1487..1503
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1529..1907
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1845..1861
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     ncRNA           2498..2619
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2647..2661
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2718..2830
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(3748..3765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3919..4507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3999..4018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4671..5462,5463..5531))
                     /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     5294..5313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5532..5636)
                     /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 attcccaata aagcctcttg ctgtttgcat
      541 ccgaatcgtg gactcgctga tccttgggag ggtctcctca gattgattga ctgcccacct
      601 cgggggtctt tcatttggag gttccaccga gatttggaga cccctgccca gggaccaccg
      661 acccccccgc cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc tgtctttgtg
      721 cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt actagttagc taactagctc
      781 tgtatctggc ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg
      841 agacgtccca gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga
      901 tcgttttgga ctctttggtg caccccccta ataggaggga tatgtggttc tggtaggaga
      961 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag
     1021 ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     1081 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     1141 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1201 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1261 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1321 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1381 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1441 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1501 cactccttct ctaggcgccc ccatatggcc atatgagatc ttatatgggg cacccccgcc
     1561 ccttgtaaac ttccctgacc ctgacatgac aagagttact aacagcccct ctctccaagc
     1621 tcacttacag gctctctact tagtccagca cgaagtctgg agacctctgg cggcagccta
     1681 ccaagaacaa ctggaccgac cggccggtgg tacctcaccc ttaccgagtc ggcgacacag
     1741 tgtgggtccg ccgacaccag actaagaacc tagaacctcg ctggaaagga ccttacacag
     1801 tcctgctgac cacccccacc gccctcaaag tagacggcat cgcagcttgg atacacgccg
     1861 cccacgtgaa ggctgccgac cccgggggtg gaccatcctc tagactgccg gatctagcta
     1921 gttaattaac tccgtggatc cggtcgtgcg gccgcacgga aacgcgtgcc accatgaacc
     1981 ccgccatctc tgtggccctg ctgctgtctg tgctgcaggt gtccagaggc cagaaagtga
     2041 ccagcctgac cgcctgcctc gtgaaccaga acctgagact ggactgcaga cacgagaaca
     2101 acaccaagga caacagcatc cagcacgagt tctccctgac ccgcgagaag agaaaacacg
     2161 tgctgagcgg caccctgggc atccccgagc acacctacag atccagagtg accctgagca
     2221 accagcccta catcaaggtg ctgaccctgg ccaacttcac cacaaaggac gagggcgact
     2281 acttctgcga actgcaggtg tcaggcgcca accccatgag cagcaacaag agcatcagcg
     2341 tgtacagaga caagctcgtg aagtgcggcg gcatcagcct gctggtgcag aacacctctt
     2401 ggatgctgtt gctgctgctg agcctgagcc tgctgcaggc cctggacttc atctctctgt
     2461 aggcatgccg cgaccctgca ggtttctgaa gctttgtttg aatgaggctt cagtacttta
     2521 cagaatcgtt gcctgcacat cttggaaaca cttgctggga ttacttcttc aggttaaccc
     2581 aacagaaggc tcgagaaggt atattgctgt tgacagtgag cgccctcttc atctacggag
     2641 tcattagtga agccacagat gtaatgactc cgtagatgaa gaggatgcct actgcctcgg
     2701 acttcaaggg gctagaattc aaggggctac tttaggagca attatcttgt ttactaaaac
     2761 tgaatacctt gctatctctt tgatacattt ttacaaagct gaattaaaat ggtataaatt
     2821 aaatcacttt atcgattgca ttgggtcgac cgggccgcca gcacagtggt cgatcgaggg
     2881 ggggcccgcg attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt
     2941 tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata gaataaaaga
     3001 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     3061 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     3121 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     3181 agcagttcct gccccggctc agggccaaga acagttggaa cagcagaata tgggccaaac
     3241 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     3301 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     3361 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     3421 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     3481 agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     3541 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     3601 gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta atttggtttt ttttcttaag
     3661 tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3721 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3841 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3901 ggccgcgttg ctggcgtttt tccataggct ccgcacccct gacgagcatc acaaaaatcg
     3961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4081 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4321 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4561 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgcg
     4621 gccggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4681 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4741 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4801 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4861 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4921 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4981 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5041 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5101 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5161 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5221 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5281 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5341 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5401 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5461 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5521 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5581 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5641 tttc
//
