LOCUS       sequence-363881-        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector, hygromycin resistance hTFAM expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  AUTHORS   Kozhukhar N, Spadafora D, Rodriguez YAR, Alexeyev MF
  TITLE     A Method for In Situ Reverse Genetic Analysis of Proteins Involved 
            mtDNA Replication.
  JOURNAL   Cells. 2022 Jul 11;11(14). pii: cells11142168. doi: 
            10.3390/cells11142168.
  PUBMED    35883613
REFERENCE   2  (bases 1 to 6955)
  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: "Cells."; 
            date: "2022-07-11"; volume: "11(14). pii: cells11142168. doi"; 
            pages: " 10.3390/cells11142168"
COMMENT     Sequence Label: pMA5132
FEATURES             Location/Qualifiers
     source          1..6955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1403..1412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2211..2227
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2211..2227
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2240..2260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2258..2555
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2425..2560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2565..2574
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2571..3599
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MDKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     misc_feature    3612..4200
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3665..3685)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4083..4094)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5044..5061)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5215..5803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5295..5314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5967..6758,6759..6827))
                     /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     6590..6609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6828..6932)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatcaat tcgccagcac
     1381 agtggtcggg ctgcaggaat tcgccaccat ggcgtttctc cgaagcatgt ggggcgtgct
     1441 gagtgccctg ggaaggtctg gagcagagct gtgcaccggc tgtggaagtc gactgcgctc
     1501 ccccttcagt tttgtgtatt taccgaggtg gttttcatct gtcttggcaa gttgtccaaa
     1561 gaaacctgta agttcttacc ttcgattttc taaagaacaa ctacccatat ttaaagctca
     1621 gaacccagat gcaaaaacta cagaactaat tagaagaatt gcccagcgtt ggagggaact
     1681 tcctgattca aagaaaaaaa tatatcaaga tgcttatagg gcggagtggc aggtatataa
     1741 agaagagata agcagattta aagaacagct aactccaagt cagattatgt ctttggaaaa
     1801 agaaatcatg gacaaacatt taaaaaggaa agctatgaca aaaaaaaaag agttaacact
     1861 gcttggaaaa ccaaaaagac ctcgttcagc ttataacgtt tatgtagctg aaagattcca
     1921 agaagctaag ggtgattcac cgcaggaaaa gctgaagact gtaaaggaaa actggaaaaa
     1981 tctgtctgac tctgaaaagg aattatatat tcagcatgct aaagaggacg aaactcgtta
     2041 tcataatgaa atgaagtctt gggaagaaca aatgattgaa gttggacgaa aggatcttct
     2101 acgtcgcaca ataaagaaac aacgaaaata tggtgctgag gagtgttaac tcgagggggg
     2161 gcccggtacc gaggacgcgt caattcgagc tccaccgcgg tggcggccgc tctagaacta
     2221 gtggatcccc cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2281 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2341 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2401 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2461 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2521 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggccacc atggataaga
     2581 aacctgaact gacagcaact tctgttgaga agtttctcat tgaaaaattt gattctgttt
     2641 ctgatctcat gcagctgtct gaaggtgaag aaagcagagc cttttctttt gatgttggag
     2701 gaagaggtta tgttctgagg gtcaattctt gtgctgatgg tttttacaaa gacagatatg
     2761 tttacagaca ctttgcctct gctgctctgc caattccaga agttctggac attggagaat
     2821 tttctgaatc tctcacctac tgcatcagca gaagagcaca aggagtcact ctccaggatc
     2881 tccctgaaac tgagctgcca gctgttctgc aacctgttgc tgaagcaatg gatgccattg
     2941 cagcagctga tctgagccaa acctctggat ttggtccttt tggtccccaa ggcattggtc
     3001 agtacaccac ttggagggat ttcatttgtg ccattgctga tcctcatgtc tatcactggc
     3061 agactgtgat ggatgacaca gtttctgctt ctgttgctca ggcactggat gaactcatgc
     3121 tgtgggcaga agattgtcct gaagtcagac acctggtcca tgctgatttt ggaagcaaca
     3181 atgttctgac agacaatggc agaatcactg cagtcattga ctggtctgaa gccatgtttg
     3241 gagattctca atatgaggtt gccaacattt ttttttggag accttggctg gcttgcatgg
     3301 aacaacaaac aagatatttt gaaagaagac acccagaact ggctggttcc cccagactga
     3361 gagcctacat gctcagaatt ggcctggacc aactgtatca atctctggtt gatggaaact
     3421 ttgatgatgc tgcttgggca caaggaagat gtgatgccat tgtgaggtct ggtgctggaa
     3481 ctgttggaag aactcaaatt gcaagaaggt ctgctgctgt ttggactgat ggatgtgttg
     3541 aagttctggc tgactctgga aacaggagac cctccacaag acccagagcc aaggaatgat
     3601 ccggaatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3661 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3721 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3781 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3841 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3901 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3961 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     4021 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4081 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4141 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     4201 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     4261 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     4321 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     4381 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     4441 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4501 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     4561 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     4621 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     4681 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     4741 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     4801 tctgagtgat tgactmcmmk wmascskskk kcwktyrcat scrrcwtgtg gtctcgctgt
     4861 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca
     4921 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt
     4981 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5041 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5101 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5161 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5401 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5641 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5701 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5761 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5821 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5881 attatcaaaa aggatcttca cctagatcct tttgcggccg gccgcaaatc aatctaaagt
     5941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
