LOCUS       sequence-10634-e        6027 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 6027)
  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 6027)
  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: pBY2887
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             105..125
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             132..152
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(171..173,174..176,177..887)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(171..173,174..176,177..887)
                     /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(216..237)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(477..496)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     824..845
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             945..989
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1032..1061
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1151..1170)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1216..1271
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1270..1289)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      1359..1406
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     1419..2124
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           1507..1879
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(1536..2124)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(2133..2166)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(2334..2921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2413..2432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2934..3725,3726..3794))
                     /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     3557..3576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     4037..4864
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4037..4161
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4199..4804)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        4932..5235
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5237..5439
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5390..5410
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5642..5664
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5726..5745
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     5787..5806
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5787..5805
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5806..5830
                     /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        5846..5955
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             5979..6002
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6009..6023
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 cgcgggcccg ggatccaccg gttactacca tggctccacg accatccaac aaacgtctcc
       61 agcagtccgg tatgccacga ccctccaaca agcgtcttca acaggagaac ctttactttc
      121 aaggtcaatt ggaaaatctc tatttccagg gaccaccagc gccaccacag atggtgagca
      181 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      241 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      301 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      361 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      421 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      481 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      541 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      601 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
      661 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
      721 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
      781 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      841 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggat tatgatattc
      901 caactactgc aagcgagaat ttggcagcac agggtgaggg tggtggcctg aacgacatct
      961 tcgaggccca gaagatcgag tggcacgagt aagcggccgc aggagctccg ggcttctcct
     1021 caatttccgc tcatcaccac catcatcacc atcaccacca ctaagtgatt aacctcaggt
     1081 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     1141 ctgagatcga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     1201 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     1261 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     1321 atttggttta gagtttggca acatatgccc atatgtaact agcataaccc cttggggcct
     1381 ctaaacgggt cttgaggggt tttttgctga aagcatgcgg aggaaattct ccttgaagtt
     1441 tccctggtgt tcaaagtaaa ggagtttgca ccagacgcac ctctgttcac tggtccggcg
     1501 tattaaaaca cgatacattg ttattagtac atttattaag cgctagattc tgtgcgttgt
     1561 tgatttacag acaattgttg tacgtatttt aataattcat taaatttata atctttaggg
     1621 tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt
     1681 aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc
     1741 gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct
     1801 tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg
     1861 acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat
     1921 tgactcgacg taaacacgtt aaatagagct tggacatatt taacatcggg cgtgttagct
     1981 ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac
     2041 gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa
     2101 tttgtagttg agctttttgg aattgcgatc gcataacttc gtatagcata cattatacga
     2161 agttataagc tcggaacgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2221 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2281 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2341 aggctccgcc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2401 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2461 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2521 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2581 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2641 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2701 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2761 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2821 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2881 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgttaccaa
     2941 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3001 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3061 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3121 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3181 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3241 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3301 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3361 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3421 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3481 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3541 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3601 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3661 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3721 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3781 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3841 ctcatgtccg cgcgtttcct gcatctttta atcaaatccc aagatgtgta taaacgcgcc
     3901 ggtatgtaca ggaagaggtt tatactaaac tgttacattg caaacgtggt ttcgtgtgcc
     3961 aagtgtgaaa accgatgttt aatcaaggct ctgacgcatt tctacaacca cgactccaag
     4021 tgtgtgggtg aagtcatgca tcttttaatc aaatcccaag atgtgtataa accaccaaac
     4081 tgccaaaaaa tgaaaactgt cgacaagctc tgtccgtttg ctggcaactg caagggtctc
     4141 aatcctattt gtaattattg aataataaaa caattataaa tgtcaaattt gttttttatt
     4201 aacgatacaa accaaacgca acaagaacat ttgtagtatt atctataatt gaaaacgcgt
     4261 agttataatc gctgaggtaa tatttaaaat cattttcaaa tgattcacag ttaatttgcg
     4321 acaatataat tttattttca cataaactag acgccttgtc gtcttcttct tcgtattcct
     4381 tctctttttc atttttctct tcataaaaat taacatagtt attatcgtat ccatatatgt
     4441 atctatcgta tagagtaaat tttttgttgt cataaatata tatgtctttt ttaatggggt
     4501 gtatagtacc gctgcgcata gtttttctgt aatttacaac agtgctattt tctggtagtt
     4561 cttcggagtg tgttgcttta attattaaat ttatataatc aatgaatttg ggatcgtcgg
     4621 ttttgtacaa tatgttgccg gcatagtacg cagcttcttc tagttcaatt acaccatttt
     4681 ttagcagcac cggattaaca taactttcca aaatgttgta cgaaccgtta aacaaaaaca
     4741 gttcacctcc cttttctata ctattgtctg cgagcagttg tttgttgtta aaaataacag
     4801 ccattgtaat gagacgcaca aactaatatc acaaactgga aatgtctatc aatatatagt
     4861 tgctgatggc cggcctatta atagtaatca attacggggt cattagttca tagcccatat
     4921 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4981 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5041 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5101 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     5161 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     5221 atcgctatta ccatgctgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
     5281 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     5341 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
     5401 ggtaggcgtg tacggtggga ggtctatata agcagacgtc gtttagtgaa ccgtcagatc
     5461 actagatgct ttattgcggt agtttatcac agttaaattg ctaacgccag tctcgaactt
     5521 aacgtgcaga agttggtcgt gaggcactgg gcaggtaagt atcgggccct ttgtgcgggg
     5581 ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg
     5641 gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc
     5701 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
     5761 ggcaaagaat tggatcggac cgaaattaat acgactcact ataggggaat tgtgagcgga
     5821 taacaattcc ccggagttaa tccgggacct ttaattcaac ccaacacaat atattatagt
     5881 taaataagaa ttattatcaa atcatttgta tattaattaa aatactatac tgtaaattac
     5941 attttattta caatcaaagg agatatacca tggcaagctg gagccacccg cagttcgaaa
     6001 agggtgcaga tgacgacgac aaggtac
//
