LOCUS       sequence-271177-        6515 bp    DNA     circular SYN 14-APR-2025
DEFINITION  retrovirus construct for stable expressing mEGFP tagged protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6515)
  AUTHORS   Lazarou M, Sliter DA, Kane LA, Sarraf SA, Wang C, Burman JL, Sideris
            DP, Fogel AI, Youle RJ
  TITLE     The ubiquitin kinase PINK1 recruits autophagy receptors to induce 
            mitophagy.
  JOURNAL   Nature. 2015 Aug 20;524(7565):309-14. doi: 10.1038/nature14893. Epub
            2015 Aug 12.
  PUBMED    26266977
REFERENCE   2  (bases 1 to 6515)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature14893"; journalName: "Nature"; date: "2015-08-20- 
            20"; volume: "524"; issue: "7565"; pages: "309-14"
COMMENT     Sequence Label: pBMN-mEGFP-C1
FEATURES             Location/Qualifiers
     source          1..6515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  148..915
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     LTR             1309..1778
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1843..2200
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2265..2681
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2611..2633
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2649..2665
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2691..3065
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     3003..3019
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      3068..3077
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3074..3076,3077..3079,3080..3790)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3119..3140)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3380..3399)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3727..3748
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3791..3856
                     /label=MCS
                     /note="multiple cloning site"
     LTR             3922..4515
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4646..4663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4817..5405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4897..4916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5579..6370,6371..6439))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6202..6221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     join(6507..6515,1..10)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atcacgaggc cctttcgtct tcaagaacag ctttgctctt aggagtttcc taatacatcc
       61 caaactcaaa tatataaagc atttgacttg ttctatgccc tagggggcgg ggggaagcta
      121 agccagcttt ttttaacatt taaaatgggt aatgactcca acttattgat agtgttttat
      181 gttcagataa tgcccgatga ctttgtcatg cagctccacc gattttgaga acgacagcga
      241 cttccgtccc agccgtgcca ggtgctgcct cagattcagg ttatgccgct caattcgctg
      301 cgtatatcgc ttgctgatta cgtgcagctt tcccttcagg cgggattcat acagcggcca
      361 gccatccgtc atccatatca ccacgtcaaa gggtgacagc aggctcataa gacgccccag
      421 cgtcgccata gtgcgttcac cgaatacgtg cgcaacaacc gtcttccgga gactgtcata
      481 cgcgtaaaac agccagcgct ggcgcgattt agccccgaca tagccccact gttcgtccat
      541 ttccgcgcag acgatgacgt cactgcccgg ctgtatgcgc gaggttaccg actgcggcct
      601 gagtttttta agtgacgtaa aatcgtgttg aggccaacgc ccataatgcg ggctgttgcc
      661 cggcatccaa cgccattcat ggccatatca atgattttct ggtgcgtacc gggttgagaa
      721 gcggtgtaag tgaactgcag ttgccatgtt ttacggcagt gagagcagag atagcgctga
      781 tgtccggcgg tgcttttgcc gttacgcacc accccgtcag tagctgaaca ggagggacag
      841 ctgatagaaa cagaagccac tggagcacct caaaaacacc atcatacact aaatcagtaa
      901 gttggcagca tcaccttaat tccattttaa atgcacagat gtttttattt cataagggtt
      961 tcaatgtgca tgaatgctgc aatattcctg ttaccaaagc tagtataaat aaaaatagat
     1021 aaacgtggaa attacttaga gtttctgtca ttaacgtttc cttcctcagt tgacaacata
     1081 aatgcgctgc tgagaagcca gtttgcatct gtcaggatca atttcccatt atgccagtca
     1141 tattaattac tagtcaatta gttgattttt atttttgaca tatacatgtg aaagacccca
     1201 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     1261 aactgagaat agaaaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     1321 caaagcggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     1381 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     1441 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     1501 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     1561 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     1621 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     1681 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     1741 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
     1801 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
     1861 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
     1921 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tcggaacacc
     1981 cggccgcaac cctgggagac gtcccaggga cttcgggggc cgtttttgtg gcccgacctg
     2041 agtccaaaaa tcccgatcgt tttggactct ttggtgcacc ccccttagag gagggatatg
     2101 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     2161 cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc
     2221 tctgtctgac tgtgtttctg tatttgtctg aaaatatggg cccgggccag actgttacca
     2281 ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt
     2341 cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta
     2401 acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca
     2461 aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg
     2521 aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc
     2581 ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc
     2641 gatcctccct ttatccagcc ctcactcctt ctctaggcgc ccccatatgg ccatatgaga
     2701 tcttatatgg ggcacccccg ccccttgtaa acttccctga ccctgacatg acaagagtta
     2761 ctaacagccc ctctctccaa gctcacttac aggctctcta cttagtccag cacgaagtct
     2821 ggagacctct ggcggcagcc taccaagaac aactggaccg accggtggta cctcaccctt
     2881 accgagtcgg cgacacagtg tgggtccgcc gacaccagac taagaaccta gaacctcgct
     2941 ggaaaggacc ttacacagtc ctgctgacca cccccaccgc cctcaaagta gacggcatcg
     3001 cagcttggat acacgccgcc cacgtgaagg ctgccgaccc cgggggtgga ccatcctcta
     3061 gactgccgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3121 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3181 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     3241 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     3301 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3361 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3421 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3481 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     3541 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     3601 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3661 gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg
     3721 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3781 gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac
     3841 cgcgggcccg ggatccaccg gatctagata actgatcacg ataaaataaa agattttatt
     3901 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     3961 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc
     4021 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     4081 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
     4141 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     4201 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     4261 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     4321 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     4381 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     4441 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     4501 cgggggtctt tcattcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt
     4561 atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4621 ttgcgtattg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4681 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4741 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4801 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4861 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4921 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4981 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5041 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5101 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5161 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5221 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5281 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5341 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5401 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5461 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5521 aaaaatgaag tttgcgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5641 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5701 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5761 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5821 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5881 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5941 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6001 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6061 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6121 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6181 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6241 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6301 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6361 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6421 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6481 attgtctcat gacattaacc tataaaaata ggcgt
//
