LOCUS       sequence-237983-        8600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible FLAG (C-terminal tag) SCAF4 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8600)
  AUTHORS   Gregersen LH, Mitter R, Ugalde AP, Nojima T, Proudfoot NJ, Agami R, 
            Stewart A, Svejstrup JQ
  TITLE     SCAF4 and SCAF8, mRNA Anti-Terminator Proteins.
  JOURNAL   Cell. 2019 May 10. pii: S0092-8674(19)30457-X. doi: 
            10.1016/j.cell.2019.04.038.
  PUBMED    31104839
REFERENCE   2  (bases 1 to 8600)
  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: "Cell. 2019 
            May 10. pii: S0092-8674(19)30457-X. doi: 
            10.1016/j.cell.2019.04.038."
COMMENT     Sequence Label: pFRT/TO/SCAF4-FLAG
FEATURES             Location/Qualifiers
     source          1..8600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4247..4270
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4318..4335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4324..4548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4681..4700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4832..4879
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4887..5909
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4887..5909
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6039..6160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6076..6095)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6130..6149
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6209..6225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6209..6225)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6222..6244)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6233..6249
                     /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(6257..6263,6264..6281,6282..6287))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6302..6323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6440..6457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6611..7199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6691..6710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7370..8161,8162..8230))
                     /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     7993..8012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8231..8335)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8410..8429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt
      781 ggtggaattc tgcagatcgg gatccatgga cgccgtcaac gccttcaacc aggagctctt
      841 ttcgcttatg gatatgaaac ctcccatctc tagagccaag atgattctca tcactaaagc
      901 tgctattaaa gctattaagc tttataagca tgtagttcaa atagtagaaa agttcatcaa
      961 aaagtgtaaa ccagaataca aggttccggg attatatgta attgactcaa ttgtgcgaca
     1021 gtctcgtcat cagtttggaa ctgataaaga tgtttttggg ccaagattct ctaaaaacat
     1081 aactgccaca ttccaatatt tatatctttg tccatctgaa gataagagta aaatagttcg
     1141 tgtgctgaac ctttggcaaa aaaatggagt gttcaaaatt gaaattattc aacctctttt
     1201 ggacatggca gcgggaacca gtaatgcagc cccagtagca gaaaatgtta ccaataatga
     1261 aggctcacct ccacctccag taaaagtttc ttctgaacct cccacacaag ccactccaaa
     1321 ctccgtccca gctgtaccac agttgcccag ctctgatgct tttgctgctg tggctcagct
     1381 gtttcagaca actcaaggcc aacagcttca gcagatcctt cagacttttc aacagcctcc
     1441 aaaaccacag tctcctgccc ttgacaatgc tgtgatggct caggttcagg ctatcacagc
     1501 tcagttaaag acaactccta cacaaccatc tgaacaaaaa gctgctttcc ccccacctga
     1561 acagaaaact gcatttgaca agaagttgct tgatagattt gactatgatg atgagccaga
     1621 agctgtggaa gaatcaaaga aagaggatac cactgccgtc accacgacag cacctgctgc
     1681 cgcagtaccc cctgcaccca ccgccaccgt gcctgctgct gctgcacccg ctgctgcctc
     1741 tcctcctcct ccacaggcac catttggctt tcctggagat ggcatgcagc agccagcata
     1801 cacacagcat caaaatatgg atcagtttca gccacgaatg atgggaatac aacaggatcc
     1861 aatgcaccat caggttccac ttcctcctaa tggacaaatg ccaggatttg gacttcttcc
     1921 tacacctcca tttcctccca tggctcagcc tgtgattcct ccaactccac cagtgcagca
     1981 gcctttccaa gcttcttttc aggcacaaaa tgaaccactt acacagaagc cgcatcagca
     2041 ggaaatggaa gtagaacaac cttgtattca agaggttaag cgacatatgt ctgataacag
     2101 aaagtcaaga tctaggtcag catccaggtc accaaaaagg aggcgatcta gatctggttc
     2161 tagatctcga aggtctcggc atcgacgttc tcgatctcgg tccagggata gacgccgaca
     2221 ttctccccga tctcgatctc aagaaagacg ggatcgagaa aaagagagag aacgtcgaca
     2281 aaaaggcctc cctcaagtga aaccggaaac tgcaagtgtt tgcagtacta ccctctgggt
     2341 ggggcagctg gacaaaagaa ctactcagca ggatgttgcc agtctcttgg aagagtttgg
     2401 tccaattgaa tcaattaata tgattcctcc caggggttgt gcctatattg ttatggttca
     2461 taggcaagat gcctatcgtg ccctgcagaa actgagccga ggaaactata aagtgaacca
     2521 gaaatccata aagattgcct gggccttaaa caaaggaata aaggcagatt ataagcagta
     2581 ttgggatgta gaacttggtg ttacttatat tccatgggac aaagtcaagc ctgaggaact
     2641 ggagagtttt tgtgaaggag gaatgttgga cagtgacaca cttaacccag attggaaagg
     2701 aattcctaag aagcctgaaa atgaagttgc tcaaaatgga ggtgctgaaa cctcacacac
     2761 agaaccagta tcacccatac ctaaaccatt acctgtgcct gtccctccta ttcctgttcc
     2821 tgcacctata acagtgccac ctccacaggt cccaccacat caaccgggtc cacctgtagt
     2881 tggtgctctc cagccgcctg ctttcacgcc tcctctggga ataccgcctc caggctttgg
     2941 tcctggtgtt cctcctcccc ctcctcctcc accatttttg cgcccaggat tcaacccaat
     3001 gcatttacca ccaggttttc tgcctcctgg acccccacct cctataactc caccagtatc
     3061 cattcctcct cctcacactc caccaataag catcccaaac tctactatcg ctggtataaa
     3121 tgaagacact acaaaagact tatctattgg aaatcccatt ccaacagtgg tgtctggggc
     3181 tagaggaaac gccgagtctg gtgacagcgt gaaaatgtat ggctctgccg tgccacctgc
     3241 tgcacccacg aatctgccca cccctcctgt aacccagcct gtttcacttc ttggcactca
     3301 aggagttgcc cctggtcctg taattggact tcaggcacca tctactggtc ttcttggcgc
     3361 ccggcccggt ctcatcccac tccagcgccc tccaggaatg cccccacctc acttacagcg
     3421 gttccctttg atgccgcccc gtcccatgcc accgcacatg atgcacagag gcccaccgcc
     3481 aggaccaggg ggctttgcga tgcctccacc tcatggaatg aaaggtccct tcccaccgca
     3541 tggccccttt gttaggcctg gtggaatgcc agggctcggg ggcccagggc caggcccagg
     3601 gggtcctgaa gacagagacg gaaggcaaca gccgccgcag cagccacagc agcagccaca
     3661 gccgcaggcg ccccagcaac cacagcagca gcagcagcag cagccaccac catcacaaca
     3721 gcctccacca acacagcagc agccacagca gtttagaaat gataacaggc agcagttcaa
     3781 ttcaggtaga gaccaagaaa ggtttggaag aagatctttt ggaaataggg tggaaaatga
     3841 ccgggaacgg tatgggaacc gtaatgatga tagagataat agtaaccgtg acaggagaga
     3901 gtggggaagg aggagccctg accgggacag gcacagagac ttggaagaga gaaatagacg
     3961 ctctagtggg catcgagaca gagagagaga ttctagagat agagagtctc gtagagagaa
     4021 ggaagaagcc cgaggaaagg aaaagcctga ggtgacagac agggcaggtg gtaacaaaac
     4081 cgttgaacct cccattagcc aagtgggaaa tgtagacact gcttcagaac ttgagaaggg
     4141 ggtgtctgag gctgcagtcc taaagccttc tgaagagtta cctgctgagg ctacctcatc
     4201 cgttgaaccc gaaaaggatt ctggctcagc agcagaggct cctcgtgact acaaagacga
     4261 tgacgacaag taagcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     4321 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4381 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4441 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4501 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     4561 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4621 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4681 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4741 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4801 aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc
     4861 tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga
     4921 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     4981 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     5041 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     5101 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     5161 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     5221 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     5281 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     5341 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     5401 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     5461 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     5521 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     5581 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     5641 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     5701 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     5761 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     5821 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     5881 ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg
     5941 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6001 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     6061 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6121 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     6181 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6241 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6301 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6361 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6421 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6481 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6541 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6601 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6661 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6721 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6781 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6841 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6901 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6961 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7021 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7081 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7141 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7201 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7261 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7321 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7381 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7441 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7501 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7561 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7621 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7681 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7741 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7801 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7861 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7921 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7981 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8041 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8101 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8161 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8221 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8281 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8341 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     8401 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     8461 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     8521 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     8581 acgggccaga tatacgcgtt
//
