LOCUS       sequence-363888-        7982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A retroviral vector encoding hygromycin resistance and PhiC31 
            recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7982)
  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 7982)
  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: pMA5136
FEATURES             Location/Qualifiers
     source          1..7982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     316..333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             463..932
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1100..1688
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1206..1217
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1615..1635
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1701..2729)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MDKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     regulatory      2726..2735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      2740..2875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(2745..3042)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     3040..3060
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3073..3089)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3073..3089)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(3160..3180)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3181..4995)
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     regulatory      4992..5001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5046..5462)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5062..5078)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5094..5116)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5663..5862
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             5927..6396
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        6427..6531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6532..6600,6601..7392)
                     /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     complement(6750..6769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7556..7982,1..162)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
       61 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
      361 ccgcctctcc ccgcgcgttg gccgattcat taatgcaact atggccattt aatgtaaata
      421 cttaagaaaa aaaaccaaat taattttgat acatgctgca tgtgaaagac ccccgctgac
      481 gggtagtcaa tcactcagag gagaccctcc caaggaacag cgagaccaca agtcggatgc
      541 aactgcaaga gggtttattg gatacacggg tacccgggcg actcagtcaa tcggaggact
      601 ggcgccccga gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga
      661 acagaagcga gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc
      721 cttggggcac cctggaaaca tctgatggtt ctctagaaac tgctgagggc tggaccgcat
      781 ctggggacca tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct
      841 gtttggccca tattcagctg ttccatctgt tcttggccct gagccggggc aggaactgct
      901 taccacagat atcctgtttg gcccatattc agctgttcca tctgttcctg accttgatct
      961 gaacttctct attctcagtt atgtattttt ccatgccttg caaaatggcg ttacttaagc
     1021 tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa
     1081 aatcttttat tttatcgatg cggggaggcg gcccaaaggg agatccgact cgtctgaggg
     1141 cgaaggcgaa gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc
     1201 gctggattga gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca
     1261 acacaggcga gcagccaagg aaaggacgat gatttccccg acaacaccac ggaattgtca
     1321 gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc
     1381 gtggcaatag ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg
     1441 ccccaaccag tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca
     1501 acgggccaca actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg
     1561 aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc
     1621 gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga
     1681 ggttgattat cgattccgga tcattccttg gctctgggtc ttgtggaggg tctcctgttt
     1741 ccagagtcag ccagaacttc aacacatcca tcagtccaaa cagcagcaga ccttcttgca
     1801 atttgagttc ttccaacagt tccagcacca gacctcacaa tggcatcaca tcttccttgt
     1861 gcccaagcag catcatcaaa gtttccatca accagagatt gatacagttg gtccaggcca
     1921 attctgagca tgtaggctct cagtctgggg gaaccagcca gttctgggtg tcttctttca
     1981 aaatatcttg tttgttgttc catgcaagcc agccaaggtc tccaaaaaaa aatgttggca
     2041 acctcatatt gagaatctcc aaacatggct tcagaccagt caatgactgc agtgattctg
     2101 ccattgtctg tcagaacatt gttgcttcca aaatcagcat ggaccaggtg tctgacttca
     2161 ggacaatctt ctgcccacag catgagttca tccagtgcct gagcaacaga agcagaaact
     2221 gtgtcatcca tcacagtctg ccagtgatag acatgaggat cagcaatggc acaaatgaaa
     2281 tccctccaag tggtgtactg accaatgcct tggggaccaa aaggaccaaa tccagaggtt
     2341 tggctcagat cagctgctgc aatggcatcc attgcttcag caacaggttg cagaacagct
     2401 ggcagctcag tttcagggag atcctggaga gtgactcctt gtgctcttct gctgatgcag
     2461 taggtgagag attcagaaaa ttctccaatg tccagaactt ctggaattgg cagagcagca
     2521 gaggcaaagt gtctgtaaac atatctgtct ttgtaaaaac catcagcaca agaattgacc
     2581 ctcagaacat aacctcttcc tccaacatca aaagaaaagg ctctgctttc ttcaccttca
     2641 gacagctgca tgagatcaga aacagaatca aatttttcaa tgagaaactt ctcaacagaa
     2701 gttgctgtca gttcaggttt cttatccatg gtggccctag gcctccaaaa aagcctcctc
     2761 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     2821 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     2881 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     2941 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     3001 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     3061 acagggtcgg gggatccact agttctagag cggccgccac cgcggtggag ctcgaattga
     3121 cgcgtcctcg gtaccgggcc ccccctcgag aagctttcac actttccgct ttttcttagg
     3181 ggcggccacg tcctcggtgc cgtcctgggc gtcatcctcg tcgtcgtcgg tgggaggctt
     3241 ggcccaggtg atgctggctc tcttctcgat gggggtgccc tggcccctgc cggtggtgct
     3301 cttggtcacc acgatcttat ccacgaacag gcccacgaac accctcttgt cgtccacgct
     3361 ggctctgccc caccaggact tggggccggt ggggtcggcg tcggcatcct cggggaacca
     3421 ctgatccagg ggcagcttag gggcctcggc ggcctccagc tcggccagtc tttcctcggc
     3481 gccctgctgt ctcagggtca gggcggcctg ctgctttctg aagtgcttcc tgcccacagg
     3541 gccgtcatag gctccggcgg ctctgtcctc gtacagctcc tccagggcgt tcagggcgtc
     3601 ggctctctcg gccaccaggt tggccctctc gccgctcttc tcgggggcct cggtcagctt
     3661 gccgaatctt ctggcggcct cccacagcag ggccagggtc tcctcgtcgc cctcggcgtg
     3721 ccggatcttg ttgaagatcc gctcggccac gaacttgtcc agggcggcca tgctcacatt
     3781 acaggtgccc tcgtgctggc caggggcgct ggggtccacc acctttcttc tccggcaccg
     3841 gtagctgtcc ttgatgctct cctcgcctct cttgctggtc atcacggcgc cacactcaca
     3901 gtacagcttg tccatggcgc tcaggatggc ctggcctctg ctcaggccct tgcctctgcc
     3961 tctgccgtcc agccaggcct gcagctcgta ccactcggca ggctcgatga tagggccaca
     4021 gtccagctcc acaggtctca gggtgatggg gtctctctgg attctgtagc cctcgatctt
     4081 ggtggtgggg gtgccgtcgg gcttcttctt gtagatcacc tcggcggcga agccggcgat
     4141 cctagggtcc ctcaggattc tcatcacggt ggcggggtcc caggcgctgc tggcggtttt
     4201 cttgccgatg gtctcgcctc tggtgggcac ggcgtcggcg tccattctct tacacaggcc
     4261 ggtgatgctg ccggggtgga tggcggcctg gctgccgggc ttgaaaggca ggtgcttgtg
     4321 ggtcttgatc tcccgccacc accacctgat cacgtcgggc tcaaactcga aggggccggt
     4381 caggggggtg gtgctgtggg ccagcttgtt gatcaccacg ttcaccatcc tgccgttccg
     4441 ggtgatctcc ttggtctcgg acaccagctc gaagccgtag ggggccttgc cgcccacata
     4501 gccgcccagc tccctctgca ggttcttggt gtccaggatc ttggcgctct tcaggctgct
     4561 ctccttgtgg ctggcgtcca gcctcatgat caggtggatc aggtccatca cgttgccctg
     4621 tctaaagacg ccttcctggg tggacacgat ggtcacgccc agggccagca gctcggacac
     4681 gatagggatg gcgtccatca ccttcagcct gctgaaccgg gacacgtcgt acacgatgat
     4741 catgttcagc ctgccggccc tacactcgtt caggattctc tcgaactcgg gtctctcggc
     4801 ggtgccgaag gcgctggtgc caggggcctc gctgaagtgg cccacaaatc tgaatctgcc
     4861 gccgtccctc tccacctctc tctgcagatc ggcggcctta tcctcgttgg cgcttctctg
     4921 ggtggcgggg ctggcggcgc tgctgttctc tctctcccgg ctctgtctgt cgtaggctcc
     4981 ggcgtaggta tccatggtgg cgaattcctg cagcccgacc actgtgctgg cgaattgatc
     5041 cggccggcgc ctagagaagg agtgagggct ggataaaggg aggatcgagg cggggtcgaa
     5101 cgaggaggtt caagggggag agacggggcg gatggaggaa gaggaggcgg aggcttaggg
     5161 tgtacaaagg gcttgaccca gggagggggg tcaaaagcca aggcttccca ggtcacgatg
     5221 taggggacct ggtctgggtg tccatgcggg ccaggtgaaa agaccttgat cttaacctgg
     5281 gtgatgaggt ctcggttaaa ggtgccgtct cgcggccatc cgacgttaaa ggttggccat
     5341 tctgcagagc agaaggtaac ccaacgtctc ttcttgacat ctaccgactg gttgtgagcg
     5401 atccgctcga catctttcca gtgacctaag gtcaaactta agggagtggt aacagtctgg
     5461 ccctaatttt cagacaaata cagaaacaca gtcagacaga gacaacacag aacgatgctg
     5521 cagcgctgca gcagacaaga cgcgcggctt cggttccaaa ccgaaagcaa aaattcagac
     5581 ggaggcggga actgttttag gttctcgtct cctaccagaa ccacatatcc tgacggggtc
     5641 ggattccaca tcgactccct tcctcaggtc gggccacaaa aacggccccc aaagtccctg
     5701 ggacgtctcc cagggttgcg gccgggtgtt cagaactcgt cagttccacc acgggtccgc
     5761 cagatacaga gctagttagc taactagtac cgacgcaggc gcataaaatc agtcatagac
     5821 actagacaat cggacagaca cagataagtt gctggccagc ttgcctcccg gtggtgggtc
     5881 ggtggtccct gggcaggggt ctcccgatcc cggacgagcc cccaaatgaa agacccccgc
     5941 tgacgggtag tcaatcactc agaggagacc ctcccaagga acagcgagac cacaagtcgg
     6001 atgcaactgc aagagggttt attggataca cgggtacccg ggcgactcag tcaatcggag
     6061 gactggcgcc ccgagtgagg ggttgtgggc tcttttattg agctcgggga gcagaagcgc
     6121 gcgaacagaa gcgagaagcg aactgattgg ttagttcaaa taaggcacag ggtcatttca
     6181 ggtccttggg gcaccctgga aacatctgat ggttctctag aaactgctga gggctggacc
     6241 gcatctgggg accatctgtt cttggccctg agccggggca ggaactgctt accacagata
     6301 tcctgtttgg cccatattca gctgttccat ctgttcttgg ccctgagccg gggcaggaac
     6361 tgcttaccac agatatcctg tttggcccat attcaggctg caggtggcac ttttcgggga
     6421 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6481 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6541 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6601 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6661 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6721 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6781 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6841 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6901 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6961 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7021 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     7081 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7141 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7201 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7261 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7321 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7381 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttgcggccg
     7441 gccgcaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7501 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7561 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7621 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7681 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7741 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7801 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7861 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7921 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7981 ag
//
