LOCUS       sequence-363933-        6376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-excisable TFAM in Blasticidin-resistannce retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6376)
  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 6376)
  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: pMA4000
FEATURES             Location/Qualifiers
     source          1..6376
                     /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"
     protein_bind    1410..1443
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1455..1471
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1455..1471
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      1491..1500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2304..2337
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2363..2383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2368..2697
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2548..2683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2699..3097
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3110..3698
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3163..3183)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3581..3592)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3866..4335
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4465..4482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4636..5224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4716..4735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5388..6179,6180..6248))
                     /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     6011..6030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6249..6353)
                     /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 tgggccgcga taacttcgta taatgtatgc tatacgaagt
     1441 tatacgcggg ccgctctaga actagtggat cccccgggct gcaggaattc gccaccatgg
     1501 cgtttctccg aagcatgtgg ggcgtgctga gtgccctggg aaggtctgga gcagagctgt
     1561 gcaccggctg tggaagtcga ctgcgctccc ccttcagttt tgtgtattta ccgaggtggt
     1621 tttcatctgt cttggcaagt tgtccaaaga aacctgtaag ttcttacctt cgattttcta
     1681 aagaacaact acccatattt aaagctcaga acccagatgc aaaaactaca gaactaatta
     1741 gaagaattgc ccagcgttgg agggaacttc ctgattcaaa gaaaaaaata tatcaagatg
     1801 cttatagggc ggagtggcag gtatataaag aagagataag cagatttaaa gaacagctaa
     1861 ctccaagtca gattatgtct ttggaaaaag aaatcatgga caaacattta aaaaggaaag
     1921 ctatgacaaa aaaaaaagag ttaacactgc ttggaaaacc aaaaagacct cgttcagctt
     1981 ataacgttta tgtagctgaa agattccaag aagctaaggg tgattcaccg caggaaaagc
     2041 tgaagactgt aaaggaaaac tggaaaaatc tgtctgactc tgaaaaggaa ttatatattc
     2101 agcatgctaa agaggacgaa actcgttatc ataatgaaat gaagtcttgg gaagaacaaa
     2161 tgattgaagt tggacgaaag gatcttctac gtcgcacaat aaagaaacaa cgaaaatatg
     2221 gtgctgagga gtgttaactc gagggggggc ccggtaccga ggacgcgtca attcgagctc
     2281 caccgcggtg gcggcccgcg tctataactt cgtataatgt atgctatacg aagttatggg
     2341 cggcctgatc ccccgaccct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2401 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2461 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2521 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2581 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2641 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaat
     2701 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     2761 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     2821 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     2881 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     2941 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     3001 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     3061 tgaattgctg ccctctggtt atgtgtggga gggctaatcc ggaatcgata atcaacctct
     3121 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3181 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3241 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3301 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3361 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3421 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3481 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     3541 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3601 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3661 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataaaata aaagatttta
     3721 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     3781 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     3841 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     3901 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
     3961 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     4021 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     4081 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     4141 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt
     4201 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     4261 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     4321 agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat
     4381 ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4441 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4501 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4561 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5041 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5101 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5221 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5281 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc
     5341 ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5401 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5461 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5521 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5581 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5641 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5701 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5761 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5821 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5881 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5941 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6001 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6061 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6361 ccccgaaaag tgccac
//
