LOCUS       sequence-363886-        7114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A retroviral vector with G418 resistance for cloning inserts between
            PhiC31 attP and attB sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7114)
  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 7114)
  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: pMA4659
FEATURES             Location/Qualifiers
     source          1..7114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(1170..1186)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1170..1186)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     1243..1259
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1244..1260
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1285..1318
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     misc_feature    1365..1938
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1533..1550)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1760..1779
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1941..2738
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1941..2738
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1998..2017)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2608..2627
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    2807..3395
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2860..2880)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3278..3289)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3403..3420
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        3583..3990
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     primer_bind     complement(4732..4751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4851..4873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4911..4929)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4997..5101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5102..5170,5171..5962)
                     /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(5320..5339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6133..6721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6622..6641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6875..6892
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc tagtgtttgc gcctgcgtct
      661 gtactagttg gctaactaga tctgtatctg gcggtcccgc ggaagaactg acgagttcgt
      721 attcccggcc gcagcccctg ggagacgtcc cagcggcctc gggggcccgt tttgtggccc
      781 attctgtatc agttaaccta cccgagtcgg actttttgga gctccgccac tgtccgaggg
      841 gtacgtggct ttgttggggg acgagagaca gagacacttc ccgcccccgt ctgaattttt
      901 gctttcggtt ttacgccgaa accgcgccgc gcgtcttgtc tgctgcagca tcgttctgtg
      961 ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagctcgaca aagttaagta
     1021 atagtccctc tctccaagct cacttacagg cggccgggct gcaggaatta ttcttactcc
     1081 acacaggcat agctacgccc ccaactgaga gaactcaaag gttaccccag ttggggcact
     1141 acacgcgatc gaattcctgc agcccggggg atccactagt tctagagcgg ccgccaccgc
     1201 ggtggagctc gaattgacgc gtcctcggta ccgggccccc cctcgaggtc gacggtatcg
     1261 ataagcttga tcgcgtgtgg agtacgcgcc cggggagccc aagggcacgc cctggcaccc
     1321 gcaccgccaa gtacatccca agcaataact gatcccccgg ccgccccctc tcccyccccc
     1381 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     1441 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     1501 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     1561 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     1621 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     1681 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     1741 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     1801 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     1861 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggtttccct
     1921 ttgaaaaaca cgataatacc atggccattg aacaagatgg attgcacgca ggttctccgg
     1981 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2041 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2101 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     2161 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2221 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2281 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2341 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2401 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2461 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccacggc gatgcctgct
     2521 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     2581 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     2641 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     2701 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagt ccggactcag atccagcgct
     2761 gataacccag cctccaccac aaacaaggat gccggtaccg gccgataatc aacctctgga
     2821 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2881 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2941 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3001 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3061 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3121 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3181 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     3241 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3301 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3361 gacgagtcgg atctcccttt gggccgcctc cccgcatcgg cccaccacca ccaccaccac
     3421 atgtgaatga tccaagctta tcgataggcc taggcctatc gataggccta ggcctatcga
     3481 taggcctaac acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     3541 ggaatgaaag accccacctg taggtttggc aagctagctg cagtaacgcc attttgcaag
     3601 gcatggaaaa ataccaaacc aagaatagag aagttcagat caagggcggg tacatgaaaa
     3661 tagctaacgt tgggccaaac aggatatctg cggtgagcag tttcggcccc ggcccggggc
     3721 caagaacaga tggtcaccgc agtttcggcc ccggcccgag gccaagaaca gatggtcccc
     3781 agatatggcc caaccctcag cagtttctta agacccatca gatgtttcca ggctccccca
     3841 aggacctgaa atgaccctgc gccttatttg aattaaccaa tcagcctgct tctcgcttct
     3901 gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc acaacccctc actcggcgcg
     3961 ccagtcctcc gacagactga gtcgcccggg tacccgtgtt ctcaataaac cctcttgcag
     4021 ttgcatccga ctcgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     4081 ccgtcagcgg gggtctttca gtaggatctc gaccgatgcc cttgagagcc ttcaacccag
     4141 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     4201 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     4261 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     4321 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     4381 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     4441 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     4501 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     4561 tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg gcgatttatg
     4621 ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgcctga tgcggtattt
     4681 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     4741 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4801 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4861 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4921 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4981 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5041 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5101 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5161 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5221 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5281 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5341 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5401 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5461 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5521 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5581 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5641 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5701 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5761 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5821 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5881 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5941 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6001 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6061 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6121 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6181 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6241 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6301 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6361 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6421 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6481 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6541 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6601 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6661 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6721 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6781 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6841 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6901 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagggt
     6961 cgaggggggg cccgcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
     7021 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagatagaat
     7081 aaaagatttt atttagtctc cagaaaaagg gggg
//
