LOCUS       sequence-467300-        7727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mitochondrial roGFP2-Orp1 (retroviral 
            vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7727)
  AUTHORS   Gutscher M, Sobotta MC, Wabnitz GH, Ballikaya S, Meyer AJ, Samstag 
            Y, Dick TP
  TITLE     Proximity-based protein thiol oxidation by H2O2-scavenging 
            peroxidases.
  JOURNAL   J Biol Chem. 2009 Nov 13;284(46):31532-40. doi: 
            10.1074/jbc.M109.059246. Epub 2009 Sep 15.
  PUBMED    19755417
REFERENCE   2  (bases 1 to 7727)
  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: 
            "10.1074/jbc.M109.059246"; journalName: "J Biol Chem"; date: 
            "2009-11-13- 13"; volume: "284"; issue: "46"; pages: "31532-40"
COMMENT     Sequence Label: pLPCX mito roGFP2-Orp1
FEATURES             Location/Qualifiers
     source          1..7727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1565..2164
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1565..1584)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2061..2081
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2339..2642
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2643..2846
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2797..2817
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(3110..3112,3113..3823)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3152..3173)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3413..3432)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3760..3781
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4466..5059
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5162..5184)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5312..5345
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5405..5438
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5458..5477
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5558..5575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5729..6317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5809..5828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6488..7279,7280..7348))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7111..7130
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7349..7453)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7521..7539
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aatgatcccc cgggctgcag gtcggccgcc
     1501 acgaccggtg ccgccaccat cccctgaccc acgcccctga cccctcacaa ggagacgacc
     1561 ttccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     1621 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     1681 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     1741 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     1801 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     1861 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     1921 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     1981 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     2041 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     2101 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     2161 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc
     2221 cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgac
     2281 tctagtaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2341 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2401 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2461 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2521 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2581 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2641 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     2701 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     2761 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     2821 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     2881 ggactcagat ctcgagatgg cctccactcg tgtcctcgcc tctcgcctgg cctcccagat
     2941 ggctgcttcc gccaaggttg cccgccctgc tgtccgcgtt gctcaggtca gcaagcgcac
     3001 catccagact ggctcccccc tccagaccct caagcgcacc cagatgacct ccatcgtcaa
     3061 cgccaccacc cgccaggctt tccagaagcg cgcctactct tccggatccg tgagcaaggg
     3121 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3181 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3241 gaagttcatc tccaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3301 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     3361 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3421 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3481 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3541 ctacaactgc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3601 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3661 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcacctg
     3721 ctccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3781 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagactagtg gtggttcagg
     3841 tggtggtggt tcaggtggtg gtggttcagg tggaggagga tcaggaggag gaggatcagg
     3901 aggaggagga tcaggaggag aattcgatat ctcagaattc tataagctag cacctgttga
     3961 caagaaaggc caaccattcc ccttcgacca attaaaggga aaagtggtgc ttatcgttaa
     4021 tgttgcctcc aaatgtggat tcactcctca atacaaagaa ctagaggcct tgtacaaacg
     4081 ttataaggac gaaggattta ccatcatcgg gttcccatgc aaccagtttg gccaccaaga
     4141 acctggctct gatgaagaaa ttgcccagtt ctgccaactg aactatggcg tgactttccc
     4201 cattatgaaa aaaattgacg ttaatggtgg caatgaggac cctgtttaca agtttttgaa
     4261 gagccaaaaa tccggtatgt tgggcttgag aggtatcaaa tggaattttg aaaaattctt
     4321 agtcgataaa aagggtaaag tgtacgaaag atactcttca ctaaccaaac cttcttcgtt
     4381 gtccgaaacc atcgaagaac ttttgaaaga ggtggaatag atcgataaaa taaaagattt
     4441 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     4501 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     4561 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     4621 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     4681 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     4741 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4801 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4861 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     4921 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4981 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     5041 tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc
     5101 accgacccac caccgggagg taagctggct gcctcgcgcg tttcggtgat gacggtgaaa
     5161 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     5221 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     5281 cccagtcacg tagcgatagc ggagtgtact gataacttcg tataatgtat gctatacgaa
     5341 gttattaggt ctgaagagga gtttacgtcc agccaagcta gggccgcgat ccggaaccct
     5401 taatataact tcgtataatg tatgctatac gaagttatca gtactggctt aactatgcgg
     5461 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     5521 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5581 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5641 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5701 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5761 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5821 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5881 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5941 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6001 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6061 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6121 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6181 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6241 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6301 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6361 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6421 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6481 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6541 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6601 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6661 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6721 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6781 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6841 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6901 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6961 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7021 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7081 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     7141 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7201 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7261 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7321 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7381 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7441 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7501 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattgc
     7561 tagcaattgc tagcaattgc tagcaattca taccagatca ccgaaaactg tcctccaaat
     7621 gtgtccccct cacactccca aattcgcggg cttctgcctc ttagaccact ctaccctatt
     7681 ccccacactc accggagcca aagccgcggc ccttccgttt ctttgct
//
