LOCUS       sequence-43217-e        4975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4975)
  AUTHORS   Schaffer U, Schlosser A, Muller KM, Schafer A, Katava N, Baumeister 
            R, Schulze E
  TITLE     SnAvi--a new tandem tag for high-affinity protein-complex 
            purification.
  JOURNAL   Nucleic Acids Res. 2010 Apr;38(6):e91. doi: 10.1093/nar/gkp1178. 
            Epub 2010 Jan 4.
  PUBMED    20047968
REFERENCE   2  (bases 1 to 4975)
  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: "Nucleic 
            Acids Res."; date: "2010-04"; volume: "38(6):e91. doi"; pages: " 
            10.1093/nar/gkp1178. Epub 2010 Jan 4"
COMMENT     Sequence Label: pBY2727
FEATURES             Location/Qualifiers
     source          1..4975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..81
                     /label=MCS
                     /note="multiple cloning site"
     CDS             164..184
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             191..211
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(230..232,233..235,236..946)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(230..232,233..235,236..946)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(275..296)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(536..555)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     883..904
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1004..1048
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    1173..1294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1210..1229)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1264..1283
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1301..1756)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1438..1459)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1650..1669
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1783..1887
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1889..2246
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1889..2080
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1912..1932)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2097..2232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2281..3075
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2281..3075
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2335..2354)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2945..2964
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3263..3282)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3307..3354
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3683..4271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4172..4191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4446..4749
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4750..4953
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4904..4924
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gctagcgcta ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
       61 gcgggcccgg gatccaccgg ttactaccat ggctccacga ccatccaaca aacgtctcca
      121 gcagtccggt atgccacgac cctccaacaa gcgtcttcaa caggagaacc tttactttca
      181 aggtcaattg gaaaatctct atttccaggg accaccagcg ccaccacaga tggtgagcaa
      241 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      301 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      361 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      421 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
      481 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      541 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      601 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
      661 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
      721 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
      781 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
      841 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
      901 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggatt atgatattcc
      961 aactactgca agcgagaatt tggcagcaca gggtgagggt ggtggcctga acgacatctt
     1021 cgaggcccag aagatcgagt ggcacgagta agcggccgcg actctagatc ataatcagcc
     1081 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1141 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1201 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1261 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     1321 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1381 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1441 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1501 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1561 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1621 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1681 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1741 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1861 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     1921 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1981 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2041 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2101 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2161 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2221 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     2281 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2341 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     2401 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2461 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     2521 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2581 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2641 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2701 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2761 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     2821 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2881 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2941 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3001 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3061 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3121 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3181 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     3241 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3301 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     3361 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     3421 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     3481 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     3541 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3601 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3661 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3721 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3781 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3841 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3901 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3961 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4021 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4081 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4141 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4201 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4261 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4321 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     4381 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     4441 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     4501 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     4561 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     4621 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     4681 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     4741 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4801 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4861 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4921 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcc
//
