LOCUS       sequence-370207-        6518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is the Lazarou lab's standard retrovirus vector backbone for 
            expression of BFP-tag fusions onto the N-terminus of cloned genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6518)
  AUTHORS   Nguyen TN, Sawa-Makarska J, Khuu G, Lam WK, Adriaenssens E, 
            Fracchiolla D, Shoebridge S, Bernklau D, Padman BS, Skulsuppaisarn 
            M, Lindblom RSJ, Martens S, Lazarou M
  TITLE     Unconventional initiation of PINK1/Parkin mitophagy by Optineurin.
  JOURNAL   Mol Cell. 2023 May 18;83(10):1693-1709.e9. doi: 
            10.1016/j.molcel.2023.04.021.
  PUBMED    37207627
REFERENCE   2  (bases 1 to 6518)
  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"; doi: 
            "10.1016/j.molcel.2023.04"; journalName: "Mol Cell"; date: 
            "2023-05-18- 18"; volume: "83"; issue: "10"; pages: "1693-1709.e9"
COMMENT     Sequence Label: pBMN BFP
FEATURES             Location/Qualifiers
     source          1..6518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(401..418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(572..1160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(652..671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1324..2115,2116..2184))
                     /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     1947..1966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2185..2289)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  2434..3201
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     LTR             3603..4072
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4137..4494
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4559..4975
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4905..4927
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4943..4959
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4985..5359
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5297..5313
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             5368..6066
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    6067..6132
                     /label=MCS
                     /note="multiple cloning site"
     LTR             join(6320..6518,1..271)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
       61 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
      121 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
      181 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
      241 tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt
      301 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
      361 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      421 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      481 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      541 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      601 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      661 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      721 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      781 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      841 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      901 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      961 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1021 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1081 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1141 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1201 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1261 agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1321 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1381 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1441 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1501 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1561 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1621 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1681 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1741 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1801 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1861 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1921 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1981 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2041 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2101 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2161 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2221 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2281 ggttccgcgc acatttcccc gaaaagtgcc accagctttg ctcttaggag tttcctaata
     2341 catcccaaac tcaaatatat aaagcatttg acttgttcta tgccctaggg ggcgggggga
     2401 agctaagcca gcttttttta acatttaaaa tgtggtaatg actccaactt attgatagtg
     2461 ttttatgttc agataatgcc cgatgacttt gtcatgcagc tccaccgatt ttgagaacga
     2521 cagcgacttc cgtcccagcc gtgccaggtg ctgcctcaga ttcaggttat gccgctcaat
     2581 tcgctgcgta tatcgcttgc tgattacgtg cagctttccc ttcaggcggg attcatacag
     2641 cggccagcca tccgtcatcc atatcaccac gtcaaagggt gacagcaggc tcataagacg
     2701 ccccagcgtc gccatagtgc gttcaccgaa tacgtgcgca acaaccgtct tccggagact
     2761 gtcatacgcg taaaacagcc agcgctggcg cgatttagcc ccgacatagc cccactgttc
     2821 gtccatttcc gcgcagacga tgacgtcact gcccggctgt atgcgcgagg ttaccgactg
     2881 cggcctgagt tttttaagtg acgtaaaatc gtgttgaggc caacgcccat aatgcgggct
     2941 gttgcccggc atccaacgcc attcatggcc atatcaatga ttttctggtg cgtaccgggt
     3001 tgagaagcgg tgtaagtgaa ctgcagttgc catgttttac ggcagtgaga gcagagatag
     3061 cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcagtagc tgaacaggag
     3121 ggacagctga tagaaacaga agccactgga gcacctcaaa aacaccatca tacactaaat
     3181 cagtaagttg gcagcatcac cttaaaatgt taattccatt ttaaatgcac agatgttttt
     3241 atttcataag ggtttcaatg tgcatgaatg ctgcaatatt cctgttacca aagctagtat
     3301 aaataaaaat agataaacgt ggaaattact tagagtttct gtcattaacg tttccttcct
     3361 cagttgacaa cataaatgcg ctgctgagaa gccagtttgc atctgtcagg atcaatttcc
     3421 cattatgcca gtcatattaa ttactagtca attagttgat ttttattttt gacatataca
     3481 tgtgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     3541 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
     3601 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3661 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     3721 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     3781 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     3841 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     3901 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
     3961 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     4021 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
     4081 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
     4141 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
     4201 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     4261 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
     4321 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg cacccccctt
     4381 agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     4441 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     4501 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata tgggcccggg
     4561 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     4621 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     4681 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     4741 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     4801 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     4861 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     4921 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat
     4981 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga
     5041 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt
     5101 ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt
     5161 ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa
     5221 cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa
     5281 agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg
     5341 tggaccatcc tctagactgc cgccaccatg agcgagctga ttaaggagaa catgcacatg
     5401 aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa
     5461 ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc
     5521 cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac
     5581 cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag
     5641 agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
     5701 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct
     5761 gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac
     5821 ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc
     5881 gcaaacatca agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc
     5941 gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc
     6001 gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag
     6061 ctgaattccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg
     6121 ggcccgggat ccaccggatc tagataactg atcacgataa aataaaagat tttatttagt
     6181 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     6241 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg
     6301 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     6361 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     6421 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     6481 agccctcagc agtttctaga gaaccatcag atgtttcc
//
