LOCUS       sequence-492421-        6417 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  Retrovirus expression Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6417)
  AUTHORS   Kabaria SR, Bae Y, Ehmann ME, Lende-Dorn BA, Beitz AM, Peterman EL, 
            Love KS, Ploessl DS, Galloway KE
  TITLE     Programmable promoter editing for precise control of transgene 
            expression.
  JOURNAL   Nat Biotechnol. 2025 Oct 13. doi: 10.1038/s41587-025-02854-y.
  PUBMED    41083901
REFERENCE   2  (bases 1 to 6417)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2025 Oct 13. doi: 10.1038/s41587-025-02854-y."
COMMENT     Sequence Label: pMXs-iCre-WPRE_pKG3028
FEATURES             Location/Qualifiers
     source          1..6417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(100..204)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             234..827
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    890..1247
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1306..1722
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1652..1674
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1690..1706
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1732..2106
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2044..2060
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    2157..2181
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2236..2255
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2306..2315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2318..2338
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3426..3446)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3844..3855)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4134..4603
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4733..4750)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4904..5492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4984..5003)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5656..6417,1..30,31..99))
                     /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     6279..6298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
       61 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      121 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      181 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg cataatgaaa
      241 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
      301 aatacataac tgagaataga aaagttcaga tcaaggtcag gaacagatgg aacagctgaa
      361 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
      421 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
      481 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
      541 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
      601 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
      661 agcccacaac ccctcactcg gcgcgccagt cctccgattg actgagtcgc ccgggtaccc
      721 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
      781 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc
      841 gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag
      901 caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc
      961 ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg
     1021 gaacacccgg ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc
     1081 cgacctgagt ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc ctaataggag
     1141 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
     1201 ttgctttcgg tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg
     1261 tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc
     1321 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
     1381 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
     1441 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
     1501 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
     1561 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
     1621 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
     1681 cgatcctccc tttatccagc cctcactcct tctctaggcg cccccatatg gccatatgag
     1741 atcttatatg gggcaccccc gccccttgta aacttccctg accctgacat gacaagagtt
     1801 actaacagcc cctctctcca agctcactta caggctctct acttagtcca gcacgaagtc
     1861 tggagacctc tggcggcagc ctaccaagaa caactggacc gaccggtggt acctcaccct
     1921 taccgagtcg gcgacacagt gtgggtccgc cgacaccaga ctaagaacct agaacctcgc
     1981 tggaaaggac cttacacagt cctgctgacc acccccaccg ccctcaaagt agacggcatc
     2041 gcagcttgga tacacgccgc ccacgtgaag gctgccgacc ccgggggtgg accatcctct
     2101 agactgccgg atctagctag ttaattaagg atcccagtgt ggtggtacgg gaaatcacaa
     2161 gtttgtacaa aaaagcaggc tccaccatgg gaaccaattc agtcgactgg atccggtacc
     2221 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     2281 tgattaattc gagcgaacgc gtgccgccac catggtgccc aagaagaaga ggaaagtctc
     2341 caacctgctg actgtgcacc aaaacctgcc tgccctccct gtggatgcca cctctgatga
     2401 agtcaggaag aacctgatgg acatgttcag ggacaggcag gccttctctg aacacacctg
     2461 gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga acaacaggaa
     2521 atggttccct gctgaacctg aggatgtgag ggactacctc ctgtacctgc aagccagagg
     2581 cctggctgtg aagaccatcc aacagcacct gggccagctc aacatgctgc acaggagatc
     2641 tggcctgcct cgcccttctg actccaatgc tgtgtccctg gtgatgagga gaatcagaaa
     2701 ggagaatgtg gatgctgggg agagagccaa gcaggccctg gcctttgaac gcactgactt
     2761 tgaccaagtc agatccctga tggagaactc tgacagatgc caggacatca ggaacctggc
     2821 cttcctgggc attgcctaca acaccctgct gcgcattgcc gaaattgcca gaatcagagt
     2881 gaaggacatc tcccgcaccg atggtgggag aatgctgatc cacattggca ggaccaagac
     2941 cctggtgtcc acagctggtg tggagaaggc cctgtccctg ggggttacca agctggtgga
     3001 gagatggatc tctgtgtctg gtgtggctga tgaccccaac aactacctgt tctgccgggt
     3061 cagaaagaat ggtgtggctg ccccttctgc cacctcccaa ctgtccaccc gggccctgga
     3121 agggatcttt gaggccaccc accgcctgat ctatggtgcc aaggatgact ctgggcagag
     3181 atacctggcc tggtctggcc actctgccag agtgggtgct gccagggaca tggccagggc
     3241 tggtgtgtcc atccctgaaa tcatgcaggc tggtggctgg accaatgtga acattgtgat
     3301 gaactacatc agaaacctgg actctgagac tggggccatg gtgaggctgc tcgaggatgg
     3361 ggactgatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3421 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3481 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3541 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3601 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3661 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3721 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3781 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3841 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3901 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgccagcaca
     3961 gtggtcgacg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     4021 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     4081 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata
     4141 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     4201 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     4261 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     4321 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     4381 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     4441 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     4501 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     4561 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa
     4621 aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa
     4681 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4741 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4801 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4861 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4921 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4981 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5041 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5101 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5161 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5221 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5281 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5341 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5401 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5461 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5521 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5581 ggatcttcac ctagatcctt ttgcggccgg ccgcaaatca atctaaagta tatatgagta
     5641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagc
//
