LOCUS       sequence-322021-        6098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for negative control of knockdown expressing a 
            non-target miR against Renilla luciferase and expression of a CD90.2
            surface marker under control of the CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6098)
  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 6098)
  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-CAG-CD90.2-Rluc_miR
FEATURES             Location/Qualifiers
     source          1..6098
                     /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"
     enhancer        1923..2302
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2305..2580
                     /label=chicken beta-actin promoter
     ncRNA           3343..3464
                     /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           3492..3506
                     /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           3563..3675
                     /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(4202..4219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4373..4961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4453..4472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5125..5916,5917..5985))
                     /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     5748..5767
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5986..6090)
                     /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 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     1981 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2041 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2101 gactttccat tgacgtcaat gggtggacta tttacggtaa actgcccact tggcagtaca
     2161 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2221 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2281 attagtcatc gctattacca tgggtcgagg tgagccccac gttctgcttc actctcccca
     2341 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     2401 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgrggggcgg
     2461 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
     2521 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
     2581 ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg ccgcccgccc
     2641 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     2701 ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt ttctgtggct gcgtgaaagc
     2761 cttaaagggc tccgggaggg ggatccggtc gtgcggccgc acggaaacgc gtgccaccat
     2821 gaaccccgcc atctctgtgg ccctgctgct gtctgtgctg caggtgtcca gaggccagaa
     2881 agtgaccagc ctgaccgcct gcctcgtgaa ccagaacctg agactggact gcagacacga
     2941 gaacaacacc aaggacaaca gcatccagca cgagttctcc ctgacccgcg agaagagaaa
     3001 acacgtgctg agcggcaccc tgggcatccc cgagcacacc tacagatcca gagtgaccct
     3061 gagcaaccag ccctacatca aggtgctgac cctggccaac ttcaccacaa aggacgaggg
     3121 cgactacttc tgcgaactgc aggtgtcagg cgccaacccc atgagcagca acaagagcat
     3181 cagcgtgtac agagacaagc tcgtgaagtg cggcggcatc agcctgctgg tgcagaacac
     3241 ctcttggatg ctgttgctgc tgctgagcct gagcctgctg caggccctgg acttcatctc
     3301 tctgtaggca tgccgcgacc ctgcaggttt ctgaagcttt gtttgaatga ggcttcagta
     3361 ctttacagaa tcgttgcctg cacatcttgg aaacacttgc tgggattact tcttcaggtt
     3421 aacccaacag aaggctcgag aaggtatatt gctgttgaca gtgagcgcag gaattataat
     3481 gcttatctat agtgaagcca cagatgtata gataagcatt ataattccta tgcctactgc
     3541 ctcggacttc aaggggctag aattcaaggg gctactttag gagcaattat cttgtttact
     3601 aaaactgaat accttgctat ctctttgata catttttaca aagctgaatt aaaatggtat
     3661 aaattaaatc actttatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     3721 aatgaaagac cccacctgta ggtttggcaa gctagctaga gaaccatcag atgtttccag
     3781 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     3841 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     3901 ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc
     3961 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     4021 attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt gtcgagcagc
     4081 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc
     4141 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4201 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4261 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4321 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4381 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4441 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4501 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4561 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4621 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4681 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4741 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4801 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4861 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4921 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4981 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5041 tatcaaaaag gatcttcacc tagatccttt tgcggccggc cgcaaatcaa tctaaagtat
     5101 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5161 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5221 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5281 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5341 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5401 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5461 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5521 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5581 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5641 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5701 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5761 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5821 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5881 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5941 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6001 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6061 ttagaaaaat aaacaaatag gggttccgcg cacatttc
//
