LOCUS       sequence-196915-        5589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Epitope tag c-terminus with SBP-Flag moiety.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5589)
  AUTHORS   Hsiao JJ, Ng BH, Smits MM, Wang J, Jasavala RJ, Martinez HD, Lee J, 
            Alston JJ, Misonou H, Trimmer JS, Wright ME
  TITLE     Androgen receptor and chemokine receptors 4 and 7 form a signaling 
            axis to regulate CXCL12-dependent cellular motility.
  JOURNAL   BMC Cancer. 2015 Mar 31;15:204. doi: 10.1186/s12885-015-1201-5.
  PUBMED    25884570
REFERENCE   2  (bases 1 to 5589)
  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: "BMC 
            Cancer."; date: "2015-03-31"; volume: "15:204. doi"; pages: " 
            10.1186/s12885-015-1201-5"
COMMENT     Sequence Label: pcDNA3-neo-Strep_Flag_Cterm
FEATURES             Location/Qualifiers
     source          1..5589
                     /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(166..184)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(167..184)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(204..221)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    210..434
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      480..908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(567..586)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     777..798
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(917..937)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        922..1251
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1102..1237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1164..1183
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1318..2112
                     /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(1372..1391)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1982..2001
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2286..2407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2323..2342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2377..2396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2456..2472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2456..2472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2469..2491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2480..2496
                     /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(2504..2510,2511..2528,2529..2534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2549..2570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2687..2704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2858..3446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2938..2957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3617..4408,4409..4477))
                     /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     4240..4259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4478..4582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4657..4676)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4848..5227
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5228..5431
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5382..5402
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5476..5495
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5476..5494
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atggacgaga agaccaccgg ctggagaggc ggccacgtgg tcgagggact ggccggtgag
       61 ctcgaacagc tacgggcccg ccttgagcac cacccccagg ggcagcgcga gccgggcggt
      121 atggactaca aggacgacga ctgatgagac tctagagggc cctattctat agtgtcacct
      181 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
      241 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
      301 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
      361 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
      421 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
      481 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
      541 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      601 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
      661 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
      721 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
      781 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
      841 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
      901 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
      961 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     1021 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     1081 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     1141 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     1201 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     1261 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     1321 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     1381 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     1441 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     1501 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     1561 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     1621 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     1681 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     1741 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     1801 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     1861 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     1921 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     1981 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     2041 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     2101 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     2161 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     2221 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     2281 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2341 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2401 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     2461 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2521 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2581 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2641 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2701 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2761 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2821 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2881 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2941 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3001 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3061 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3121 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3181 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3241 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3301 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3361 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3421 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3481 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3601 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3661 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3721 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3781 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3841 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3901 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3961 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4021 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4081 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4141 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4201 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4261 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4321 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4381 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4441 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4501 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4561 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     4621 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     4681 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     4741 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     4801 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     4861 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     4921 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     4981 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     5041 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     5101 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     5161 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     5221 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     5281 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     5341 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     5401 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     5461 ctggcttatc gaaattaata cgactcacta tagggagacc caagcttggt accgagctcg
     5521 gatccactag taacggccgc cagtgtgctg gaattctgca gatatccatc acactggcgg
     5581 ccgctcgag
//
