LOCUS       sequence-364432-        6764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mtDNA destruction/production of rho-0 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  AUTHORS   Spadafora D, Kozhukhar N, Chouljenko VN, Kousoulas KG, Alexeyev MF
  TITLE     Methods for Efficient Elimination of Mitochondrial DNA from Cultured
            Cells.
  JOURNAL   PLoS One. 2016 May 2;11(5):e0154684. doi: 
            10.1371/journal.pone.0154684. eCollection 2016.
  PUBMED    27136098
REFERENCE   2  (bases 1 to 6764)
  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: "PLoS One.";
            date: "2016-05-2"; volume: "11(5):e0154684. doi"; pages: " 
            10.1371/journal.pone.0154684. eCollection 2016"
COMMENT     Sequence Label: pMA4008
FEATURES             Location/Qualifiers
     source          1..6764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        678..782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(783..851,852..1643)
                     /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     complement(1001..1020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1814..2402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2303..2322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2556..2573
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2936..3160
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3149..3166
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3186..3204
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3186..3203
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      4557..4566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4632..4651)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4633..4651)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4696..4899)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4725..4745)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4900..5279
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    5512..5633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5523..5542)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5577..5596
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5634..6347,6348..6350,6351..6353))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5637..6347,6348..6350,6351..6353))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5679..5700)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6028..6047
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6287..6308
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      6350..6359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6369..6630)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 ccgccacact tgcaacatca cacttccgcc acactactac gtcacccgcc ccgttcccac
       61 gccccgcgcc acgtcacaaa ctccaccccc tcattatcat attggcttca atccaaaata
      121 aggtatattc atcaataata taccttattt tggattgaag ccaatatgat aatgaggggg
      181 tggagtttgt ggcgcggggc gtgggaacgg ggcgggtgac gtaggtttta gggcggagta
      241 acttgtatgt gttgggaatt gtagttttct taaaatggga agtgacgtaa cgtgggaaaa
      301 cggaagtgac gatttgagga agttgtgggt tttttggctt tcgtttctgg gcgtaggttc
      361 gcgtgcggtt ttctgggtgt tttttgtgga ctttaaccgt tacgtcattt tttagtccta
      421 tatatactcg ctctgcactt ggcccttttt tacactgtga ctgattgagc tggtgccgtg
      481 tcgagtggtg tttttttaat aggttttctt ttttactggt aaggctgact gttatggctg
      541 ccgctgtgga agcgctgtat gttgttctgg agcgggaggg tgctattttg cctagagtga
      601 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
      661 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      721 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      781 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      841 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      901 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      961 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1021 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1081 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1141 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1201 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1261 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1321 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1381 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1441 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1501 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1561 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1621 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1681 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1741 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1801 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1861 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1921 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1981 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2041 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2101 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2161 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2221 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2281 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2341 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2401 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2461 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2521 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2581 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcaggg
     2641 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     2701 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     2761 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     2821 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc gaagttccta
     2881 tactctagta agaataggaa cttcggatcg atcctggttc tttccgcctc agaagccata
     2941 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
     3001 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
     3061 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
     3121 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgagct
     3181 ctagcattta ggtgacacta tagaataggg ccctctagat gcatgctcga gtcagcgaga
     3241 cgacctcccc gtcgtcggtg atgaggacgt tgtttcggca gcagcagggc cgggccccgg
     3301 agaacgagag gcccatagct cggcgagcgt gtcgtcgaac gccaggcggc tgcttcgctg
     3361 gatggcctta tagatctccg gatcgatgcg gacgggggta atgatcaggg cgatcggaac
     3421 ggcctggttc gggagaatgg acgccttgct gggtcctgcg gccccgagag ccccggcgcc
     3481 gtcctccagg cggaacgtta cgccctcctc cgcgctggtg cggtgcctgc cgataaacgt
     3541 caccagatgc gggtgggggg ggcagtcggg gaagtggctg tcgagcacgt agccctgcac
     3601 caagatctgc ttaaagttcg ggtgacgggg gttcgcgaag acgggctcgc ggcggaccag
     3661 atccccggag ctccaggaca cgggggagat ggtgtggcgt ccgaggtcgg gggcgccaaa
     3721 cagaagcacc tccgagacaa cgccgctatt taactccacc aaggcccgat ccgcggcgga
     3781 gcaccgcctt ttttcgcccg aggcgtgggc ctctgaccag gcctggtctt gcgtgacgag
     3841 agcctcctcc gggccgggga cgcgcccggg cgcgaagtat cgcacgctgg gcttcgggat
     3901 cgaccggata aatgcccgga acgcctccgg ggaccggtgt gtcatcaagt cctcgtacgc
     3961 ggaggccgtg gggtcgctgg ggtccatggg gtcgaaagcg tacttggccc ggcatttgac
     4021 ctcgtaaaag gccagggggg tcttggggac tggggccagg tagccgtgaa tgtcccgagg
     4081 acagacgaga atatccaggg acgccccgac catccccgtg tgaccgtcca tgaggacccc
     4141 acacgtatgc acgttctctt cggtgaggtc gctgggttcg tggaagataa agcgccgcgt
     4201 gtcggcgccg gcctcgccgc cgtcgtccgc gcggcccacg cagtagcgaa acagcaggct
     4261 tcgggccgtc ggctcgttca cccgcccgaa catcaccgcc gaagactgta catccggccg
     4321 caggctggcg ttgtgcttca gccactgggg cgagaaacac ggaccctggg ggccccagcg
     4381 gagggtggat gcggtcgtga ggccccgccg gagcagggcc catagctggc agtcggcctg
     4441 gttttgcgtg gccgcctcgt aaaaccccat gaggggccgg ggcgccacgg cgtccgcggc
     4501 ggccgggggc ccgcggcgcg tcaggcgcca taggtgccgg ccgagtccgc ggtccaccat
     4561 ggtggcgaat tccagcacac tggcggccgt tactagtgga tcgatccgag ctcggtacca
     4621 agcttgggtc tccctatagt gagtcgtatt aatttcgata agccagtaag cagtgggttc
     4681 tctagttagc cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     4741 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     4801 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     4861 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     4921 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     4981 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     5041 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     5101 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     5161 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     5221 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca
     5281 acgcgtatat ctggcccgta catcgctaga gctgactata ataataaaac gccaactttg
     5341 acccggaacg cggaaaacac ctgagaaaaa cacctgggcg agtctccacg taaacggtca
     5401 aagtccccgc ggccctagac aaatattacg cgctatgagt aacacaaaat tattcagatt
     5461 tcacttcctc ttattcagtt ttcccgcgaa aatgggggta tacagacatg ataagataca
     5521 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5581 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttttacttg
     5641 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     5701 tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg
     5761 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     5821 tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc
     5881 tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     5941 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     6001 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     6061 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     6121 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc
     6181 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg
     6241 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     6301 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg
     6361 agcttggagg tgcacaccaa tgtggtgaat ggtcaaatgg cgtttattgt atcgagctag
     6421 gcacttaaat acaatatctc tgcaatgcgg aattcagtgg ttcgtccaat ccatgtcaga
     6481 cccgtctgtt gccttcctaa taaggcacga tcgtaccacc ttacttccac caatcggcat
     6541 gcacggtgct ttttctctcc ttgtaaggca tgttgctaac tcatcgttac catgttgcaa
     6601 gactacaaga gtattgcata agactacatt cccccaaatc ttactcggtt acgcccaaat
     6661 ttactacaac atccgcctaa aaccgcgcga aaattgtcac ttcctgtgta caccggcgca
     6721 caccaaaaac gtcacttttg ccacatccgt cgcttacatg tgtt
//
