LOCUS       sequence-325481-        6541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Gfi1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6541)
  AUTHORS   Menendez L, Trecek T, Gopalakrishnan S, Tao L, Markowitz AL, Yu HV, 
            Wang X, Llamas J, Huang C, Lee J, Kalluri R, Ichida J, Segil N
  TITLE     Generation of inner ear hair cells by direct lineage conversion of 
            primary somatic cells.
  JOURNAL   Elife. 2020 Jun 30;9. pii: 55249. doi: 10.7554/eLife.55249.
  PUBMED    32602462
REFERENCE   2  (bases 1 to 6541)
  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: "Elife."; 
            date: "2020-06-30"; volume: "9. pii: 55249. doi"; pages: " 
            10.7554/eLife.55249"
COMMENT     Sequence Label: pMXs-Gfi1
FEATURES             Location/Qualifiers
     source          1..6541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..358
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             417..833
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     763..785
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     801..817
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    843..1217
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1155..1171
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    1260..1284
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2560..2584)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2601..3189
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2654..2674)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3072..3083)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3369..3838
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3968..3985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4139..4727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4219..4238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4891..5682,5683..5751))
                     /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     5514..5533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5752..5856)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5886..6479
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat
       61 gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
      121 tgacgagttc ggaacacccg gccgcaaccc tgggagacgt cccagggact tcgggggccg
      181 tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt tggactcttt ggtgcacccc
      241 cctaatagga gggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc
      301 cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt gtctgctgca
      361 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
      421 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
      481 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
      541 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
      601 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
      661 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
      721 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
      781 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcccccatat
      841 ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct gaccctgaca
      901 tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc tacttagtcc
      961 agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac cgaccggtgg
     1021 tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg ccgacaccag actaagaacc
     1081 tagaacctcg ctggaaagga ccttacacag tcctgctgac cacccccacc gccctcaaag
     1141 tagacggcat cgcagcttgg atacacgccg cccacgtgaa ggctgccgac cccgggggtg
     1201 gaccatcctc tagactgccg gatctagcta gttaatcgac ggtatcgata agctatcaaa
     1261 caagtttgta caaaaaagca ggctgccacc atgccgcgct catttctcgt caaaagcaag
     1321 aaggctcaca gctaccacca gccgcgctcc ccaggaccag actattccct ccgtttagag
     1381 aatgtaccgg cgcctagccg agcagacagc acttcaaatg caggcggggc gaaggcggag
     1441 ccccgggacc gtttgtcccc cgaatcgcag ctgaccgaag ccccagacag agcctccgca
     1501 tccccagaca gctgcgaagg cagcgtctgc gaacggagct cggagtttga ggacttctgg
     1561 aggcccccgt caccctccgc gtctccagcc tcggagaagt caatgtgccc atcgctggac
     1621 gaagcccagc ccttccccct gcctttcaaa ccgtactcat ggagcggcct ggcgggttct
     1681 gacctgcggc acctggtgca gagctaccga ccgtgtgggg ccctggagcg tggcgctggc
     1741 ctgggcctct tctgtgaacc cgccccggag cctggccacc cggccgcgct gtacggcccg
     1801 aagcgggctg ccggcggcgc gggggccggg gcgccaggga gctgcagcgc aggggccggt
     1861 gccaccgctg gccctggcct agggctctac ggcgacttcg ggtctgcggc agccgggctg
     1921 tatgagaggc ccacggcagc ggcgggcttg ctgtaccccg agcgtggcca cgggctgcac
     1981 gcagacaagg gcgctggcgt caaggtggag tcggagctgc tgtgcacccg cctgctgctg
     2041 ggcggcggct cctacaagtg catcaagtgc agcaaggtgt tctccacgcc gcacgggctc
     2101 gaggtgcacg tgcgcaggtc ccacagcggt accagaccct ttgcctgcga gatgtgcggc
     2161 aagaccttcg ggcacgcggt gagcctggag cagcacaaag ccgtgcactc gcaggaacgg
     2221 agctttgact gtaagatctg tgggaagagc ttcaagaggt catccacact gtccacacac
     2281 ctgcttatcc actcagacac tcggccctac ccctgtcagt actgtggcaa gaggttccac
     2341 cagaagtcag acatgaagaa acacactttc atccacactg gtgagaagcc tcacaagtgc
     2401 caggtgtgcg gcaaggcatt cagccagagc tccaacctca tcacccacag ccgcaaacac
     2461 acaggcttca agcccttcgg ctgcgacctc tgtgggaagg gtttccagag gaaggtggac
     2521 ctccgaaggc accgggagac gcagcatggg ctcaaatgaa cccagctttc ttgtacaaag
     2581 tggttcgata gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2641 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2701 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2761 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2821 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2881 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2941 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     3001 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3061 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3121 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3181 cctccccgca tcgataccgt cgacgataaa ataaaagatt ttatttagtc tccagaaaaa
     3241 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     3301 caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga
     3361 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     3421 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     3481 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
     3541 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     3601 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     3661 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     3721 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     3781 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca
     3841 tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat
     3901 agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3961 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4021 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4561 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4801 tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccggccgca aatcaatcta
     4861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4921 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4981 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5041 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5101 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5161 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5221 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5281 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5341 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5401 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5461 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5521 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5581 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5641 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5701 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5761 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5821 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5881 tgcataatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     5941 aaggcatgga aaaatacata actgagaata gaaaagttca gatcaaggtc aggaacagat
     6001 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     6061 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     6121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
     6181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     6241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     6301 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc
     6361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     6421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
     6481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
     6541 t
//
