LOCUS       sequence-109630-        7970 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral expression of Flag-tagged murine transcription factor 
            Scl.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7970)
  AUTHORS   Batta K, Florkowska M, Kouskoff V, Lacaud G
  TITLE     Direct reprogramming of murine fibroblasts to hematopoietic 
            progenitor cells.
  JOURNAL   Cell Rep. 2014 Dec 11;9(5):1871-84. doi: 
            10.1016/j.celrep.2014.11.002. Epub 2014 Nov 26.
  PUBMED    25466247
REFERENCE   2  (bases 1 to 7970)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2014.11.002"; journalName: "Cell Rep"; date: 
            "2014-12-11- 11"; volume: "9"; issue: "5"; pages: "1871-84"
COMMENT     Sequence Label: Flag-mScl
FEATURES             Location/Qualifiers
     source          1..7970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(132..200,201..992)
                     /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     complement(350..369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1163..1751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1652..1671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1905..1922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2039..2060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2075..2080,2081..2098,2099..2105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2129
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2118..2140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2137..2153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2137..2153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2172..2192
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2174..2192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             2399..2656
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2700..2825
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3318..3551
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3735..3779
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4041..4158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4197..4408
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4417..4426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4426..4449
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             6711..6944
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7016..7137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7053..7072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7107..7126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7156..7291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7323..7342)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7324..7342)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7352..7369)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7352..7368)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7361..7383)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7513..7968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7600..7619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7810..7831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgga
     2401 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     2461 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     2521 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     2581 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     2641 tggaaaatct ctagcagtgg cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc
     2701 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     2761 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     2821 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     2881 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     2941 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     3001 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     3061 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     3121 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     3181 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     3241 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     3301 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     3361 atgggcgcag cctcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     3421 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     3481 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     3541 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     3601 tggaatgcta gttggagtaa taaatctctg gaacagattg gaatcacacg acctggatgg
     3661 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     3721 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     3781 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     3841 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     3901 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     3961 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     4021 acggatctcg acggtatcgg ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     4081 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     4141 acaaaaattc aaaattttat cgtttaaacg gcgcgccggc tccggtgccc gtcagtgggc
     4201 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
     4261 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     4321 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     4381 tcgcaacggg tttgccgcca gaacacaggt cccggggcca ccatggatta caaggatgac
     4441 gacgataaga cggagcggcc gccgagcgag gcggcacgca gtgacccgca actagaggga
     4501 caggacgcgg ccgaggcccg catggccccc ccgcacctag tcctgctcaa cggcgtcgcc
     4561 aaggagacga gccgcgcagc cccggctgag ccccccgtca tcgagctagg agcgcgcagc
     4621 ggcgcggggg gcggccctgc cagtgggggc ggtgccgcga gggacttaaa gggccgcgac
     4681 gcagtagcag ccgaagctcg ccttcgggtg cccaccaccg agctgtgcag acctcccgga
     4741 cccgccccgg cgcccgcgcc cgcctcggct cctgcagagc tgcctggaga cggccgcatg
     4801 gtgcagctga gcccgcccgc gctggcagcc cctgccggcc ccggccgagc gctgctctat
     4861 agccttagcc agccgctcgc ctcactaggc agtgggttct ttggggaacc ggatgccttc
     4921 cccatgttca ccaacaacaa ccgggtgaag aggaggccct ccccatatga gatggagatt
     4981 tctgatggtc ctcacaccaa agtagtgcgg cgcatcttca ccaacagccg ggaacgatgg
     5041 aggcagcaga atgtgaatgg ggcatttgct gagctcagaa agctgatccc cacccaccca
     5101 ccagacaaga aactaagcaa gaatgagatc ctccgccttg ccatgaagta catcaatttc
     5161 ctggccaagt tactcaatga ccaggaggag gaaggcaccc agcgtgccaa gcctggcaag
     5221 gaccccgtgg tgggagctgg tgggggtggg gcagggggtg gcatcccccc tgaagacctt
     5281 ctacaggacg tgctttcccc caactccagc tgtggcagct ctctggatgg ggcagccagc
     5341 ccggacagtt acacagagga gccaacaccc aagcacactt cccgcagcct ccatcctgcc
     5401 ctgctgcctg ccgctgatgg ggctggcccc cggtgagaat tcggccacgc gtccgcggac
     5461 tagtctcgag ttaattaagc tagcctagtg ccatttgttc agtggttcgt agggctttcc
     5521 cccactgttt ggctttcagt tatatggatg atgtggtatt gggggccaag tctgtacagc
     5581 atcttgagtc cctttttacc gctgttacca attttctttt gtctttgggt atacatttaa
     5641 accctaacaa aacaaagaga tggggttact ctctaaattt tatgggttat gtcattggat
     5701 gttatgggtc cttgccacaa gaacacatca tacaaaaaat caaagaatgt tttagaaaac
     5761 ttcctattaa caggcctatt gattggaaag tatgtcaacg aattgtgggt cttttgggtt
     5821 ttgctgcccc ttttacacaa tgtggttatc ctgcgttgat gcctttgtat gcatgtattc
     5881 aatctaagca ggctttcact ttctcgccaa cttacaaggc ctttctgtgt aaacaatacc
     5941 tgaaccttta ccccgttgcc cggcaacggc cacctctgtg ccaagtgttt gctgacgcaa
     6001 cccccactgg ctggggcttg gtcatgggcc atcagcgcat gcgtggaacc ttttcggctc
     6061 ctctgccgat ccatactgcg gaactcctag ccgcttgttt tgctcgcagc aggtctggag
     6121 caaacattat cgggactgat aactctgttg tcctatcccg caaatataca tcgtttccat
     6181 ggctgctagg ctgtgctgcc aactggatcc tgcgcgggac gtcctttgtt tacgtcccgt
     6241 cggcgctgaa tcctgcggac gacccttctc ggggtcgctt gggactctct cgtccccttc
     6301 tccgtctgcc gttccgaccg accacggggc gcacctctct ttacgcggac tccccgtctg
     6361 tgccttctca tctgccggac cgtgtgcact tcgcttcacc tctgcacgtc gcatggagac
     6421 caccgtgaac gcccaccaaa tattgcccaa ggtcttacat aagaggactc ttggactctc
     6481 agcaatgtca acgaccgacc ttgaggcata cttcaaagac tgtttgttta aagactggga
     6541 ggagttgggg gaggagatta ggttaaaggt ctttgtacta ggaggctgta ggcataaatt
     6601 ggtctgcgca ccagcaccat ggcgcaatca ctagagcggg gtacctttaa gaccaatgac
     6661 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     6721 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     6781 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6841 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     6901 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     6961 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     7021 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7081 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7141 tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     7201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     7261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcgtcgagac gtacccaatt
     7321 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     7381 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     7441 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     7501 gcgaatggcg cgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     7561 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     7621 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     7681 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     7741 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     7801 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     7861 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     7921 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttcc
//
