LOCUS       sequence-140246-        7194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes multiple copies of an epitope tag V5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7194)
  AUTHORS   Viswanathan S, Williams ME, Bloss EB, Stasevich TJ, Speer CM, Nern 
            A, Pfeiffer BD, Hooks BM, Li WP, English BP, Tian T, Henry GL, 
            Macklin JJ, Patel R, Gerfen CR, Zhuang X, Wang Y, Rubin GM, Looger 
            LL
  TITLE     High-performance probes for light and electron microscopy.
  JOURNAL   Nat Methods. 2015 Jun;12(6):568-76. doi: 10.1038/nmeth.3365. Epub 
            2015 Apr 27.
  PUBMED    25915120
REFERENCE   2  (bases 1 to 7194)
  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"; doi: 
            "10.1038/nmeth.3365"; journalName: "Nat Methods"; date: "2015-06"; 
            volume: "12"; issue: "6"; pages: "568-76"
COMMENT     Sequence Label: pCAG_smFP V5
FEATURES             Location/Qualifiers
     source          1..7194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    58..192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(82..101)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     136..155
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(198..393)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      212..347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    452..473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(488..493,494..511,512..518)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    526..542
                     /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     550..566
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     909..928
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    927..982
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1028..1047
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     1149..1165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1184..1204
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1186..1204
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1300..1411
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1400..1417
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    1444..2032
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1550..1561
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1959..1979
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(2039..2058)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2041..2173
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(2222..2263)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2273..2314)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2324..2365)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2579..2620)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2630..2671)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2690..2731)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2741..2782)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(3314..3355)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(3365..3406)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(3416..3457)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      3457..3466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3556..3575)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          3583..4599
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(3637..3659)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4600..4877)
                     /label=chicken beta-actin promoter
     enhancer        4879..5258
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5289..5393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5394..5462,5463..6254)
                     /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(5612..5631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6425..7013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6914..6933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7167..7184
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat
       61 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
      121 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
      181 aataaacaag ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
      241 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
      301 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
      361 ggcgggacta tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg
      421 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      481 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      541 aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg
      601 gatctccata agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat
      661 ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa
      721 caaaatataa aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca
      781 gcagggggct gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc
      841 agcatatggg catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga
      901 tttgccctcc catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca
      961 atgaaaataa atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc
     1021 ccataatttt tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc
     1081 acaccagcca ccaccttctg ataggcagcc tgcacctgag gagtgaatta gcggataaca
     1141 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     1201 aagggaacaa aagctgggct attgtcttcc caatcctccc ccttgctgtc ctgccccacc
     1261 ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat
     1321 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
     1381 acaacagatg gctggcaact agaaggcaca gtcgaggctg atcgtaccga gctcgaattc
     1441 caggcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg
     1501 gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg
     1561 aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc
     1621 atggaaagga cgtcagcttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag
     1681 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg
     1741 ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg
     1801 ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct
     1861 cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     1921 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     1981 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttgtcgacac
     2041 ctgtggagag aaaggcaaag tggatgtcag taagaccaat aggtgcctat cagaaacgca
     2101 agagtcttct ctgtctcgac aagcccagtt tctattggtc tccttaaacc tgtcttgtaa
     2161 ccttgatact tacctgcggc ctcgacctta gctattcatc ggtacctcga gcaccggttt
     2221 aggtactatc cagtcccagc aacgggttcg ggatgggctt gccctgctgt tgcgtactgt
     2281 cgaggcccaa cagtggattt ggtattggct ttccttgttg ctgagtgctg tcgaggccca
     2341 gcaagggatt tggtatgggc ttgcctcctt ggcccttgta cagctcgtcc attcccaacg
     2401 taatgccggc ggcggtcacg aactccaaca acaccatgtg atctcgtttt tcgtttgggt
     2461 ccttactcag aacgctctga gtcgacagat agtgattatc cggcaacagc acggggccat
     2521 ctccgatcgg cgtattctgc tgatagtggt ccgccagttg aacacttcca tcgccacctg
     2581 tcgagtcgag gcccagcaga gggttgggga ttggcttgcc ttgctgctgg gtgctatcca
     2641 ggccgagcag gggattgggt atcggtttgc caccgccttg ctggcctccg gtgctgtcga
     2701 gtcccagcaa ggggttaggg atgggctttc cctgttgctg cgtgctatcc aagcccagca
     2761 ggggattcgg gatcggcttg ccaccacctt ccacgttgtg ccgaattttg aaattcgcct
     2821 ttatgccgtt cttctgcttg tcggcagtaa tgtagacgtt atggctgtta aagttgtatt
     2881 ccagcttgtg acccagaata ttaccgtctt ccttgaaatc aattccctta agttctatcc
     2941 tgttcaccaa ggtgtctccc tcgaacttca cctcggctcg cgtcttgtaa gtaccgtcat
     3001 ctttgaaact gatcgtccgt tcctgcacgt atccctccgg catagcactc ttaaagaagt
     3061 catgctgctt catgtggtct gggtaacggc tgaaacactg gactccgcca cccagggttg
     3121 taaccaaggt cggccacggc accggcagct ttcccgtggt gcatataaac ttcagcgtca
     3181 attttccgtt ggtggcatct ccctctcctt ctccgcgcac actaaacttg tgtccgttaa
     3241 cgtcaccgtc gagctcaacc agaattggga ccacgccggt aaacagttct tcgcccttac
     3301 tcacaccttg gcccgtggag tccaggccca acagagggtt agggataggc ttaccttgtt
     3361 gctgtgtgct atccagaccg agaagtgggt ttggaatggg cttaccctgc tgctgagtac
     3421 tatcaagtcc aaggaggggg ttcggtatag gttttcccat ggtggcggcc gcaattgcgc
     3481 tagcccgggt ccggagatat ccagctggcg cgccgggccc gcggtaccga attctttgcc
     3541 aaaatgatga gacagcacaa taaccagcac gttgcccagg agctgtagga aaaagaagaa
     3601 ggcatgaaca tggttagcag aggctctaga gccgccggtc acacgccaga agccgaaccc
     3661 cgccctgccc cgtccccccc gaaggcagcc gtccccccgc ggacagcccc gaggctggag
     3721 agggagaagg ggacggcggc gcggcgacgc acgaaggccc tccccgccca tttccttcct
     3781 gccggcgccg caccgcttcg ccccgcgccc gctagagggg gtgcggcggc gcctcccaga
     3841 tttcggctcc gcacagattt gggacaaagg aagtccctgc gccctctcgc acgattacca
     3901 taaaaggcaa tggctgcggc tcgccgcgcc tcgacagccg ccggcgctcc gggggccgcc
     3961 gcgcccctcc cccgagccct ccccggcccg aggcggcccc gccccgcccg gcacccccac
     4021 ctgccgccac cccccgcccg gcacggcgag ccccgcgcca cgccccgtac ggagccccgc
     4081 acccgaagcc gggccgtgct cagcaactcg gggagggggg tgcagggggg gttgcagccc
     4141 gaccgacgcg cccacacccc ctgctcaccc ccccacgcac acaccccgca cgcagccttt
     4201 gttcccctcg cagccccccc cgcaccgcgg ggcaccgccc ccggccgcgc tcccctcgcg
     4261 cacactgcgg agcgcacaaa gccccgcgcc gcgcccgcag cgctcacagc cgccgggcag
     4321 cgcggagccg cacgcggcgc tccccacgca cacacacacg cacgcacccc ccgagccgct
     4381 ccccccgcac aaagggccct cccggagccc ctcaaggctt tcacgcagcc acagaaaaga
     4441 aacaagccgt cattaaacca agcgctaatt acagcccgga ggagaagggc cgtcccgccc
     4501 gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg aggcggcggc
     4561 ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt
     4621 tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga gcgcgccgct
     4681 ctgattggct gccgccgcac ctctccgcct cgccccgccc cgcccctcgc cccgccccgc
     4741 cccgcctggc gcgcgccccc cccccccccc cgcccccatc gctgcacaaa ataattaaaa
     4801 aataaataaa tacaaaattg ggggtgggga ggggggggag atggggagag tgaagcagaa
     4861 cgtggggctc acctcgacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     4921 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     4981 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     5041 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     5101 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     5161 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     5221 aattgattac tattaataac tagtcaataa tcaatgtcga cccaggtggc acttttcggg
     5281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6541 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
