LOCUS       sequence-196913-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Epitope tag n-terminus with SBP-Flag moiety.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  AUTHORS   Hsiao JJ, Smits MM, Ng BH, Lee J, Wright ME
  TITLE     Discovery Proteomics Identifies a Molecular Link between the 
            Coatomer Protein Complex I and Androgen Receptor-dependent 
            Transcription.
  JOURNAL   J Biol Chem. 2016 Sep 2;291(36):18818-42. doi: 
            10.1074/jbc.M116.732313. Epub 2016 Jun 30.
  PUBMED    27365400
REFERENCE   2  (bases 1 to 5570)
  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: 
            "10.1074/jbc.M116.732313"; journalName: "J Biol Chem"; date: 
            "2016-09-2- 2"; volume: "291"; issue: "36"; pages: "18818-42"
COMMENT     Sequence Label: pcDNA3-neo-Strep_Flag_Nterm
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..114
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     promoter        complement(202..220)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(203..220)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(240..257)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    246..470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      516..944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(603..622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     813..834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(953..973)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        958..1287
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1138..1273
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1200..1219
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1354..2148
                     /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(1408..1427)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2018..2037
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2322..2443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2359..2378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2413..2432
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2492..2508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2492..2508)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2505..2527)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2516..2532
                     /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(2540..2546,2547..2564,2565..2570))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2585..2606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2723..2740)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2894..3482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2974..2993)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3653..4444,4445..4513))
                     /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     4276..4295
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4514..4618)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4693..4712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4884..5263
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5264..5467
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5418..5438
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5512..5531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5512..5530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      join(5565..5570,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggacgaga agaccaccgg ctggagaggc ggccacgtgg tcgagggact ggccggtgag
       61 ctcgaacagc tacgggcccg ccttgagcac cacccccagg ggcagcgcga gccgggcggt
      121 atggactaca aggacgacga cgaattctgc agatatccat cacactggcg gccgctcgag
      181 catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc
      241 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
      301 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
      361 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
      421 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
      481 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
      541 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      601 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      661 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      721 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      781 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
      841 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
      901 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
      961 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     1021 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     1081 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     1141 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     1201 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     1261 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     1321 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     1381 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     1441 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     1501 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     1561 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     1621 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     1681 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     1741 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     1801 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     1861 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     1921 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     1981 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     2041 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     2101 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     2161 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     2221 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     2281 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     2341 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2401 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     2461 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2521 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2581 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2641 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2701 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2761 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2821 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2881 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2941 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3001 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3061 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3121 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3181 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3241 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3301 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3361 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3421 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3481 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3541 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3601 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3661 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3721 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3781 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3841 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3901 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3961 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4021 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4081 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4141 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4201 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4261 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4321 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4381 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4441 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4501 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4561 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4621 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     4681 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     4741 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     4801 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     4861 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     4921 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     4981 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5041 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5101 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     5161 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     5221 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     5281 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     5341 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     5401 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     5461 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     5521 tcactatagg gagacccaag cttggtaccg agctcggatc cgccgccacc
//
