LOCUS       sequence-248588-        6746 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of human FLI1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  AUTHORS   Palii CG, Cheng Q, Gillespie MA, Shannon P, Mazurczyk M, Napolitani 
            G, Price ND, Ranish JA, Morrissey E, Higgs DR, Brand M
  TITLE     Single-Cell Proteomics Reveal that Quantitative Changes in 
            Co-expressed Lineage-Specific Transcription Factors Determine Cell 
            Fate.
  JOURNAL   Cell Stem Cell. 2019 May 2;24(5):812-820.e5. doi: 
            10.1016/j.stem.2019.02.006. Epub 2019 Mar 14.
  PUBMED    30880026
REFERENCE   2  (bases 1 to 6746)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.stem.2019.02.006"; journalName: "Cell Stem Cell"; date: 
            "2019-05-2- 2"; volume: "24"; issue: "5"; pages: "812-820.e5"
COMMENT     Sequence Label: pcDNA3.1+/-FLI1-C‐(K)‐DYK
FEATURES             Location/Qualifiers
     source          1..6746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      15..24
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1377..1400
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1420..1437)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1426..1650
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1696..2124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1783..1802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1993..2014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2133..2153)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2138..2467
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2318..2453
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2380..2399
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2534..3328
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2588..2607)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3198..3217
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3502..3623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3539..3558)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3593..3612
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3672..3688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3672..3688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3685..3707)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3696..3712
                     /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)."
     promoter        complement(join(3720..3726,3727..3744,3745..3750))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3765..3786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3903..3920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4074..4659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4154..4173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4830..5621,5622..5690))
                     /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     5453..5472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5691..5795)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5870..5889)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6061..6440
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6441..6644
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6595..6615
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6689..6708
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6689..6707
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccgagctcgg atccgccacc atggacggga ctattaagga ggctctgtcg gtggtgagcg
       61 acgaccagtc cctctttgac tcagcgtacg gagcggcagc ccatctcccc aaggccgaca
      121 tgactgcctc ggggagtcct gactacgggc agccccacaa gatcaacccc ctcccaccac
      181 agcaggagtg gatcaatcag ccagtgaggg tcaacgtcaa gcgggagtat gaccacatga
      241 atggatccag ggagtctccg gtggactgca gcgttagcaa atgcagcaag ctggtgggcg
      301 gaggcgagtc caaccccatg aactacaaca gctatatgga cgagaagaat ggcccccctc
      361 ctcccaacat gaccaccaac gagaggagag tcatcgtccc cgcagacccc acactgtgga
      421 cacaggagca tgtgaggcaa tggctggagt gggccataaa ggagtacagc ttgatggaga
      481 tcgacacatc ctttttccag aacatggatg gcaaggaact gtgtaaaatg aacaaggagg
      541 acttcctccg cgccaccacc ctctacaaca cggaagtgct gttgtcacac ctcagttacc
      601 tcagggaaag ttcactgctg gcctataata caacctccca caccgaccaa tcctcacgat
      661 tgagtgtcaa agaagaccct tcttatgact cagtcagaag aggagcttgg ggcaataaca
      721 tgaattctgg cctcaacaaa agtcctcccc ttggaggggc acaaacgatc agtaagaata
      781 cagagcaacg gccccagcca gatccgtatc agatcctggg cccgaccagc agtcgcctag
      841 ccaaccctgg aagcgggcag atccagctgt ggcaattcct cctggagctg ctctccgaca
      901 gcgccaacgc cagctgtatc acctgggagg ggaccaacgg ggagttcaaa atgacggacc
      961 ccgatgaggt ggccaggcgc tggggcgagc ggaaaagcaa gcccaacatg aattacgaca
     1021 agctgagccg ggccctccgt tattactatg ataaaaacat tatgaccaaa gtgcacggca
     1081 aaagatatgc ttacaaattt gacttccacg gcattgccca ggctctgcag ccacatccga
     1141 ccgagtcgtc catgtacaag tacccttctg acatctccta catgccttcc taccatgccc
     1201 accagcagaa ggtgaacttt gtccctcccc atccatcctc catgcctgtc acttcctcca
     1261 gcttctttgg agccgcatca caatactgga cctcccccac ggggggaatc taccccaacc
     1321 ccaacgtccc ccgccatcct aacacccacg tgccttcaca cttaggcagc tactacgatt
     1381 acaaggatga cgacgataag tgataaaccc gctgatcagc ctcgactgtg ccttctagtt
     1441 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     1501 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1561 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     1621 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     1681 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1741 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1801 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1861 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1921 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1981 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2041 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2101 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     2161 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2221 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2281 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2341 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2401 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2461 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     2521 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2581 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2641 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2701 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2761 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2821 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2881 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2941 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3001 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3061 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3121 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3181 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3241 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3301 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3361 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3421 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3481 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3541 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3601 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     3661 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3721 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3781 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3841 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3901 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3961 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4021 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4081 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4141 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4201 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4261 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4321 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4381 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4441 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4501 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4561 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt
     4621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5821 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     5881 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     5941 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     6001 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     6061 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6121 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6181 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     6421 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     6481 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     6541 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     6601 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
     6661 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
     6721 gctagcgttt aaacttaagc ttggta
//
