LOCUS       sequence-488736-        6257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of SNAP-p53DD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6257)
  AUTHORS   Lende-Dorn BA, Atkinson JC, Bae Y, Galloway KE
  TITLE     Chemogenetic tuning reveals optimal MAPK signaling for cell-fate 
            programming
  JOURNAL   bioRxiv 2024.12.19.629350
REFERENCE   2  (bases 1 to 6257)
  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: "bioRxiv 
            2024.12.19.629350"
COMMENT     Sequence Label: pMXs-SNAP-p53DD
FEATURES             Location/Qualifiers
     source          1..6257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..133
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    222..246
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      313..322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             319..864
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     protein_bind    complement(1238..1262)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1279..1867
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1332..1352)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1750..1761)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2047..2516
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2646..2663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2817..3405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2897..2916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3569..4360,4361..4429))
                     /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     4192..4211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4430..4534)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4564..5157
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5220..5577
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5636..6052
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5982..6004
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6020..6036
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6062..6257,1..179)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
ORIGIN
        1 tcggcgacac agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag
       61 gaccttacac agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt
      121 ggatacacgc cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc
      181 cggatctagc tagttaatcg acggtatcga taagctatca aacaagtttg tacaaaaaag
      241 caggctccac catgggaacc aattcagtcg actggatccg gtactactga attcaagctt
      301 ggatcccaat gtgccaccat ggacaaagac tgcgaaatga agcgcaccac cctggatagc
      361 cctctgggca agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg
      421 ggcaaaggaa catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc
      481 ggaccagagc cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag
      541 gccatcgagg agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt
      601 acccgccagg tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac
      661 agccacctgg ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg
      721 agcggaaatc ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac
      781 gtggggggct acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac
      841 agactgggca agcctgggct gggtcctgca ggcggatcca tgactgccat ggaggagtca
      901 cagtcggata tcagcctcaa gagagcgctg cccacctgca caagcgcctc tcccccgcaa
      961 aagaaaaaac cacttgatgg agagtatttc accctcaaga tccgcgggcg taaacgcttc
     1021 gagatgttcc gggagctgaa tgaggcctta gagttaaagg atgcccatgc tacagaggag
     1081 tctggagaca gcagggctca ctccagctac ctgaagacca agaagggcca gtctacttcc
     1141 cgccataaaa aaacaatggt caagaaagtg gggcctgact cagactgaca acgttatccc
     1201 ctgattctgt ggataaccgt attacgctga tatctagacc cagctttctt gtacaaagtg
     1261 gttcgatagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1321 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1381 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     1441 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     1501 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     1561 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1621 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     1681 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     1741 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     1801 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     1861 tccccgcatc gataccgtcg acgataaaat aaaagatttt atttagtctc cagaaaaagg
     1921 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     1981 aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg
     2041 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     2101 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     2161 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
     2221 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     2281 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     2341 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgatt gactgagtcg
     2401 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     2461 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcacatg
     2521 cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag
     2581 ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2641 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2701 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2761 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2821 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2881 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2941 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3001 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3061 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3121 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3181 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3241 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3301 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3481 agattatcaa aaaggatctt cacctagatc cttttgcggc cggccgcaaa tcaatctaaa
     3541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4501 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4561 cataatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     4621 ggcatggaaa aatacataac tgagaataga aaagttcaga tcaaggtcag gaacagatgg
     4681 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4741 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     4801 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     4861 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     4921 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     4981 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgattg actgagtcgc
     5041 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     5101 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
     5161 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta
     5221 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
     5281 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
     5341 gacgagttcg gaacacccgg ccgcaaccct gggagacgtc ccagggactt cgggggccgt
     5401 ttttgtggcc cgacctgagt ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc
     5461 ctaataggag ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     5521 gtctgaattt ttgctttcgg tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag
     5581 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     5641 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     5701 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     5761 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     5821 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     5881 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     5941 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     6001 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg cccccatatg
     6061 gccatatgag atcttatatg gggcaccccc gccccttgta aacttccctg accctgacat
     6121 gacaagagtt actaacagcc cctctctcca agctcactta caggctctct acttagtcca
     6181 gcacgaagtc tggagacctc tggcggcagc ctaccaagaa caactggacc gaccggtggt
     6241 acctcaccct taccgag
//
