LOCUS       sequence-322064-        5991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector to express C-terminally mTurquoise2-tagged murine 
            Lyz2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5991)
  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 5991)
  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-Lyz2-GSG-mTurquoise
FEATURES             Location/Qualifiers
     source          1..5991
                     /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"
     CDS             join(2442..2444,2445..2447,2448..3161)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2487..2508)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2748..2767)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3095..3116
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4095..4112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4266..4854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4346..4365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5018..5809,5810..5878))
                     /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     5641..5660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5879..5983)
                     /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 accatgaaga
     1981 ctctcctgac tctgggactc ctcctgcttt ctgtcactgc tcaggccaag gtctatgaac
     2041 gttgtgagtt tgccagaact ctgaaaagga atggaatggc tggctactat ggagtcagcc
     2101 tggccgactg ggtgtgttta gctcagcacg agagcaatta taacacacga gctacaaact
     2161 acaaccgtgg agaccaaagc actgactatg ggatatttca gatcaatagc cgatactggt
     2221 gtaatgatgg caaaacccca agagctgtga atgcctgtgg gatcaattgc agtgctctgc
     2281 tgcaggatga catcactgca gccatacaat gtgcaaagag ggtggtgaga gatccccaag
     2341 gcattcgagc atgggtggca tggcgagcac actgtcaaaa ccgagatctg tcccagtata
     2401 ttcggaactg cggagtcgga tctggcgtcg acggatctgg catggtgagc aagggcgagg
     2461 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2521 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2581 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgtcct
     2641 ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2701 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2761 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2821 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactact
     2881 ttagcgacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     2941 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     3001 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     3061 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3121 ccgccgggat cactctcggc atggacgagc tgtacaagta aaagcttgag tacatcgatt
     3181 gcattgggtc gatcgaccgg gccgccagca cagtggtcga tcgagggggg gcccgcgatt
     3241 agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata
     3301 tcaccagctg aagcctatag agtacgagcc atagatagaa taaaagattt tatttagtct
     3361 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     3421 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     3481 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     3541 ccggctcagg gccaagaaca gttggaacag cagaatatgg gccaaacagg atatctgtgg
     3601 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     3661 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     3721 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     3781 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     3841 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     3901 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     3961 tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa
     4021 tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4081 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4141 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4201 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4261 gcgtttttcc ataggctccg cacccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4681 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4861 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4921 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc ggccgcaaat
     4981 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5041 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5101 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5161 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5221 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5281 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5341 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5401 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5461 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5521 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5581 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5641 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5701 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5761 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5821 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5881 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5941 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt c
//
