LOCUS       sequence-322101-        5381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral backbone for in-frame cloning of a gene of interest and a
            P2A separated CD90.2 surface marker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5381)
  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 5381)
  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-MCS-P2A-CD90.2
FEATURES             Location/Qualifiers
     source          1..5381
                     /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"
     primer_bind     complement(3485..3502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3656..4244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3736..3755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4408..5199,5200..5268))
                     /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     5031..5050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5269..5373)
                     /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 aacgcgtggc ctggcatgcc
     1981 gcgacgtcga cgccacaaac ttcagcctgc tgagacaggc tggcgacgtg gaagagaatc
     2041 ctggacctat gaaccccgcc atctctgtgg ccctgctgct gtctgtgctg caggtgtcca
     2101 gaggccagaa agtgaccagc ctgaccgcct gcctcgtgaa ccagaacctg agactggact
     2161 gcagacacga gaacaacacc aaggacaaca gcatccagca cgagttctcc ctgacccgcg
     2221 agaagagaaa acacgtgctg agcggcaccc tgggcatccc cgagcacacc tacagatcca
     2281 gagtgaccct gagcaaccag ccctacatca aggtgctgac cctggccaac ttcaccacaa
     2341 aggacgaggg cgactacttc tgcgaactgc aggtgtcagg cgccaacccc atgagcagca
     2401 acaagagcat cagcgtgtac agagacaagc tcgtgaagtg cggcggcatc agcctgctgg
     2461 tgcagaacac ctcttggatg ctgttgctgc tgctgagcct gagcctgctg caggccctgg
     2521 acttcatctc tctgtagcct gcaggtttct gaagcttgag tacatcgatt gcattgggtc
     2581 gatcgaccgg gccgccagca cagtggtcga tcgagggggg gcccgcgatt agtccaattt
     2641 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
     2701 aagcctatag agtacgagcc atagatagaa taaaagattt tatttagtct ccagaaaaag
     2761 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     2821 aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga
     2881 gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     2941 gccaagaaca gttggaacag cagaatatgg gccaaacagg atatctgtgg taagcagttc
     3001 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt
     3061 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     3121 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     3181 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
     3241 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     3301 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacatgc
     3361 agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa tggccatagt
     3421 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3481 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3541 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3601 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3661 ataggctccg cacccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3721 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3781 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3841 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3901 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3961 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4021 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4081 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4141 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4201 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4261 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4321 gattatcaaa aaggatcttc acctagatcc ttttgcggcc ggccgcaaat caatctaaag
     4381 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4441 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4501 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4561 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4621 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4681 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4741 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4801 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4861 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4921 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4981 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5041 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5101 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5161 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5221 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5281 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5341 tatttagaaa aataaacaaa taggggttcc gcgcacattt c
//
