LOCUS       sequence-491527-        6039 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  GIB-Cre Retrovirus expressing C-terminal giberellin inducible Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6039)
  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 6039)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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-GIB-Cre-C-WPRE_pKG3944
FEATURES             Location/Qualifiers
     source          1..6039
                     /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"
     CDS             338..355
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             986..1006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1020..1044)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1061..1649
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1114..1134)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1532..1543)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1829..2298
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2428..2445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2599..3187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2679..2698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3351..4142,4143..4211))
                     /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     3974..3993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4212..4316)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4346..4939
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5002..5359
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5418..5834
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5764..5786
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5802..5818
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(5844..6039,1..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 gtactactag gctgtctcgt
      301 ctcgtctcgc acgcgtgccg ccaccatgaa gcgggaccac caccatcacc atcatcagga
      361 caagaaaacc atgatgatga acgaagagga cgacggcaac ggcatggacg agctgctggc
      421 tgtgctgggc tacaaagtgc ggagcagcga gatggccgac gtggcccaga aactggaaca
      481 gctggaagtg atgatgagca acgtgcagga agatgacctg tcccagctgg ccaccgagac
      541 agtgcactac aaccccgccg agctgtacac ctggctggac tccatgctga ccgacctgaa
      601 ctccggaggg tctggctccg gatcaagtgg tggcagcggt accgtggctg atgaccccaa
      661 caactacctg ttctgccggg tcagaaagaa tggtgtggct gccccttctg ccacctccca
      721 actgtccacc cgggccctgg aagggatctt tgaggccacc caccgcctga tctatggtgc
      781 caaggatgac tctgggcaga gatacctggc ctggtctggc cactctgcca gagtgggtgc
      841 tgccagggac atggccaggg ctggtgtgtc catccctgaa atcatgcagg ctggtggctg
      901 gaccaatgtg aacattgtga tgaactacat cagaaacctg gactctgaga ctggggccat
      961 ggtgaggctg ctcgaggatg gggaccccaa gaaaaagcgg aaggtgcgct gatatctaga
     1021 cccagctttc ttgtacaaag tggttcgata gcttatcgat aatcaacctc tggattacaa
     1081 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     1141 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     1201 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     1261 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     1321 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     1381 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     1441 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     1501 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     1561 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     1621 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacgataaa ataaaagatt
     1681 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     1741 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc
     1801 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     1861 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
     1921 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     1981 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     2041 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     2101 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc
     2161 agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     2221 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     2281 gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta atttggtttt ttttcttaag
     2341 tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2401 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2461 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2521 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2581 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2641 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2701 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2761 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2821 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2881 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2941 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3001 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3061 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3121 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3181 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3241 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgcg
     3301 gccggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3361 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3421 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3481 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3541 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3601 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3661 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3721 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3781 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3841 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3901 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3961 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4021 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4081 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4141 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4201 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4261 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4321 tttccccgaa aagtgccacc tgcataatga aagaccccac ctgtaggttt ggcaagctag
     4381 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gaaaagttca
     4441 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     4501 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     4561 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     4621 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4681 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4741 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca
     4801 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4861 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4921 tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc
     4981 accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct
     5041 agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat
     5101 ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg
     5161 tcccagggac ttcgggggcc gtttttgtgg cccgacctga gtccaaaaat cccgatcgtt
     5221 ttggactctt tggtgcaccc ccctaatagg agggatatgt ggttctggta ggagacgaga
     5281 acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg
     5341 ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt
     5401 atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact
     5461 ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg
     5521 ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct
     5581 ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac
     5641 acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct
     5701 gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc
     5761 tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc
     5821 cttctctagg cgcccccata tggccatatg agatcttata tggggcaccc ccgccccttg
     5881 taaacttccc tgaccctgac atgacaagag ttactaacag cccctctctc caagctcact
     5941 tacaggctct ctacttagtc cagcacgaag tctggagacc tctggcggca gcctaccaag
     6001 aacaactgga ccgaccggtg gtacctcacc cttaccgag
//
