LOCUS       sequence-430625-        5504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  enables eukaryotic expression of N-terminally FLAG-HA-tagged fusion 
            proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  AUTHORS   Horn M, Geisen C, Cermak L, Becker B, Nakamura S, Klein C, Pagano M,
            Antebi A
  TITLE     DRE-1/FBXO11-Dependent Degradation of BLMP-1/BLIMP-1 Governs C. 
            elegans Developmental Timing and Maturation.
  JOURNAL   Dev Cell. 2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028.
  PUBMED    24613396
REFERENCE   2  (bases 1 to 5504)
  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: "Dev Cell. 
            2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028."
COMMENT     Sequence Label: FLAG-HA-pcDNA3.1-
FEATURES             Location/Qualifiers
     source          1..5504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     310..329
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     563..580
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    697..718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(733..738,739..756,757..763)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    771..787
                     /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     776..798
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     795..811
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     795..811
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    860..981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(871..890)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     925..944
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1155..1949)
                     /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(1266..1285)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1876..1895
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2016..2345)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2030..2165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2084..2103)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2330..2350
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2359..2787)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2469..2490)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2681..2700
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2833..3057
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3046..3063
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3185..3211)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3227..3250)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3250..3259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3276..3295)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3277..3295)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3340..3543)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3369..3389)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3544..3923
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4095..4114
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4189..4293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4294..4362,4363..5154)
                     /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(4512..4531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5325..5504,1..409)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
       61 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      121 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      181 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      241 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      301 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      361 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      421 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      481 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      541 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      601 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      661 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      721 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      781 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
      841 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
      901 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
      961 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1021 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1081 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     1141 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     1201 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     1261 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     1321 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     1381 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac
     1441 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     1501 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     1561 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     1621 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     1681 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     1741 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc
     1801 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     1861 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     1921 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca
     1981 gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa
     2041 aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa
     2101 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     2161 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     2221 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     2281 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     2341 cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2401 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2461 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2521 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2581 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2641 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2701 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2761 cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt
     2821 ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
     2881 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
     2941 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     3001 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     3061 aggctgatca gcggtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt
     3121 ggtggaattc tgcagatatc cagcacagtg gcggccgctc gagtctagaa cctccacctc
     3181 caccagcgta atctggaaca tcgtatgggt aacctccacc gcccccttta tcgtcatcgt
     3241 ctttgtagtc catggtggct agccagcttg ggtctcccta tagtgagtcg tattaatttc
     3301 gataagccag taagcagtgg gttctctagt tagccagaga gctctgctta tatagacctc
     3361 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     3421 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     3481 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     3541 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     3601 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     3661 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     3721 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     3781 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     3841 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     3901 ataactagtc aataatcaat gtcaacgcgt atatctggcc cgtacatcgc gaagcagcgc
     3961 aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt gccttgttgt
     4021 agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag ggagcagata
     4081 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata ggggatcggg
     4141 agatctcccg atccgtcgac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4201 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4261 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4321 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4381 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4441 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4501 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4561 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4621 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4681 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4741 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4801 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4861 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4921 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4981 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5041 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5101 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5161 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5221 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5281 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5341 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5401 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5461 ccaaatactg ttcttctagt gtagccgtag ttaggccacc actt
//
