LOCUS       sequence-181372-        6357 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of SF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  AUTHORS   Huang X, Zheng M, Wang P, Mok BW, Liu S, Lau SY, Chen P, Liu YC, Liu
            H, Chen Y, Song W, Yuen KY, Chen H
  TITLE     An NS-segment exonic splicing enhancer regulates influenza A virus 
            replication in mammalian cells.
  JOURNAL   Nat Commun. 2017 Mar 21;8:14751. doi: 10.1038/ncomms14751.
  PUBMED    28323816
REFERENCE   2  (bases 1 to 6357)
  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"; journalName: "Nat 
            Commun."; date: "2017-03-21"; volume: "8:14751. doi"; pages: " 
            10.1038/ncomms14751"
COMMENT     Sequence Label: pcDNA-FLAG-SF2
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             912..935
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1778..1795)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1784..2008
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2054..2482
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2141..2160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2351..2372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2491..2511)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2496..2826
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2677..2812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2739..2758
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2875..3900
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGGFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4030..4151
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4067..4086)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4121..4140
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4200..4216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4200..4216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4213..4235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4224..4240
                     /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(4248..4254,4255..4272,4273..4278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4293..4314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4431..4448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4602..5190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4682..4701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5361..6152,6153..6221))
                     /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     5984..6003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6222..6326)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacat ggattacaag gatgacgacg ataaggggcc ctctagactc gagcggccgc
      961 cactgtgctg gatatctgca gaattcatgt cgggaggtgg tgtgattcgt ggccccgcag
     1021 ggaacaacga ttgccgcatc tacgtgggta acttacctcc agacatccga accaaggaca
     1081 ttgaggacgt gttctacaaa tacggcgcta tccgcgacat cgacctcaag aatcgccgcg
     1141 ggggaccgcc cttcgccttc gttgagttcg aggacccgcg agacgcggaa gacgcggtgt
     1201 atggtcgcga cggctatgat tacgatgggt accgtctgcg ggtggagttt cctcgaagcg
     1261 gccgtggaac aggccgaggc ggcggcgggg gtggaggtgg cggagctccc cgaggtcgct
     1321 atggcccccc atccaggcgg tctgaaaaca gagtggttgt ctctggactg cctccaagtg
     1381 gaagttggca ggatttaaag gatcacatgc gtgaagcagg tgatgtatgt tatgctgatg
     1441 tttaccgaga tggcactggt gtcgtggagt ttgtacggaa agaagatatg acctatgcag
     1501 ttcgaaaact ggataacact aagtttagat ctcatgaggg agaaactgcc tacatccggg
     1561 ttaaagttga tgggcccaga agtccaagtt atggaagatc tcgatctcga agccgtagtc
     1621 gtagcagaag ccgtagcaga agcaacagca ggagtcgcag ttactcccca aggagaagca
     1681 gaggatcacc acgctattct ccccgtcata gcagatctcg ctctcgtaca taaggatccg
     1741 agctcggtac caagcttaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc
     1801 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1861 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     1921 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     1981 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2041 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2101 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2161 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     2221 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2281 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2341 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2401 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     2461 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2521 ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2581 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2641 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2701 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2761 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2821 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgatgaaa
     2881 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     2941 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     3001 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     3061 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggga
     3121 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     3181 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
     3241 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     3301 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     3361 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     3421 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
     3481 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     3541 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
     3601 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
     3661 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     3721 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     3781 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
     3841 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag
     3901 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3961 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4021 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4081 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4141 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4201 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4261 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4321 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4381 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     4801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5041 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5281 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5341 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5401 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5461 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5521 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5581 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5641 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5701 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5761 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5821 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5881 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5941 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6001 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6061 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6121 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6181 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6241 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6301 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
