LOCUS       sequence-321687-        5608 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 human 
            FTH1 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  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 5608)
  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-CD90.2-Cd247A_miR
FEATURES             Location/Qualifiers
     source          1..5608
                     /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"
     ncRNA           2853..2974
                     /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           3002..3016
                     /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           3073..3185
                     /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(3712..3729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3883..4471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3963..3982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4635..5426,5427..5495))
                     /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     5258..5277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5496..5600)
                     /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 acggaaacgc gtgccaccat gaaccccgcc
     2341 atctctgtgg ccctgctgct gtctgtgctg caggtgtcca gaggccagaa agtgaccagc
     2401 ctgaccgcct gcctcgtgaa ccagaacctg agactggact gcagacacga gaacaacacc
     2461 aaggacaaca gcatccagca cgagttctcc ctgacccgcg agaagagaaa acacgtgctg
     2521 agcggcaccc tgggcatccc cgagcacacc tacagatcca gagtgaccct gagcaaccag
     2581 ccctacatca aggtgctgac cctggccaac ttcaccacaa aggacgaggg cgactacttc
     2641 tgcgaactgc aggtgtcagg cgccaacccc atgagcagca acaagagcat cagcgtgtac
     2701 agagacaagc tcgtgaagtg cggcggcatc agcctgctgg tgcagaacac ctcttggatg
     2761 ctgttgctgc tgctgagcct gagcctgctg caggccctgg acttcatctc tctgtaggca
     2821 tgccgcgacc ctgcaggttt ctgaagcttt gtttgaatga ggcttcagta ctttacagaa
     2881 tcgttgcctg cacatcttgg aaacacttgc tgggattact tcttcaggtt aacccaacag
     2941 aaggctcgag aaggtatatt gctgttgaca gtgagcgccc tcttcatcta cggagtcatt
     3001 agtgaagcca cagatgtaat gactccgtag atgaagagga tgcctactgc ctcggacttc
     3061 aaggggctag aattcaaggg gctactttag gagcaattat cttgtttact aaaactgaat
     3121 accttgctat ctctttgata catttttaca aagctgaatt aaaatggtat aaattaaatc
     3181 actttatcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     3241 cccacctgta ggtttggcaa gctagctaga gaaccatcag atgtttccag ggtgccccaa
     3301 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     3361 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     3421 cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     3481 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     3541 cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt gtcgagcagc atgtatcaaa
     3601 attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat
     3661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4561 gatcttcacc tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa
     4621 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4681 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4741 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4801 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4861 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4921 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4981 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5041 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5101 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5161 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5221 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5281 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5341 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5401 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5461 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5521 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5581 aaacaaatag gggttccgcg cacatttc
//
