LOCUS       sequence-71986-e        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FX cloning mammalian cell line expression vector with CMV promoter 
            and N-terminal streptavidin binding peptide, Myc tag and 3C protease
            cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  AUTHORS   Geertsma ER, Dutzler R
  TITLE     A versatile and efficient high-throughput cloning tool for 
            structural biology.
  JOURNAL   Biochemistry. 2011 Apr 19;50(15):3272-8. Epub 2011 Mar 25.
  PUBMED    21410291
REFERENCE   2  (bases 1 to 5855)
  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: 
            "Biochemistry."; date: "2011-04-19"; volume: "50(15)"; pages: 
            "3272-8. Epub 2011 Mar 25"
COMMENT     Sequence Label: pcDXNSM3
FEATURES             Location/Qualifiers
     source          1..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    917..1079
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             1092..1205
                     /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"
     CDS             1206..1235
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1242..1265
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(1314..1619)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1416..1435)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1961..2620)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2535..2554
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2674..2680,2681..2698,2699..2704))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2729..2853)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2872..2892)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2896..2916)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2983..3000)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2989..3213
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3259..3687
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3346..3365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3556..3577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3696..3716)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3701..4030
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3881..4016
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3943..3962
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     rep_origin      complement(4103..4688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4183..4202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4859..5650,5651..5719))
                     /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     5482..5501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5720..5824)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttagcg cagaggcttg gggcagccga gcggcagcca ggccccggcc
      961 cgggcctcgg ttccagaagg gagaggagcc cgccaaggcg cgcaagagag cgggctgcct
     1021 cgcagtccga gccggagagg gagcgcgagc cgcgccggcc ccggacggcc tccgaaacca
     1081 tgggggccag catggacgag aagaccactg gttggcgagg tggacacgtt gttgaaggac
     1141 tggctgggga acttgaacaa cttcgtgcac gactggagca tcacccacaa ggtcaacgtg
     1201 aaccagaaca aaaactcatc tcagaagagg atctgggtac cttagaagtt ttgtttcaag
     1261 gtccaagtag aagagcgagc tgcagactgg ctgtgtataa gggagcctga catttatatt
     1321 ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct gagatcagcc
     1381 acttcttccc cgataacgga gaccggcaca ctggccatat cggtggtcat catgcgccag
     1441 ctttcatccc cgatatgcac caccgggtaa agttcacggg agactttatc tgacagcaga
     1501 cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat atcactctgt
     1561 acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt aaactgcatt
     1621 tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt tcaataaacc gggcgacctc
     1681 agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac gggcttcatt
     1741 ctgcatggtt gtgcttacca gaccggagat attgacatca tatatgcctt gagcaactga
     1801 tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct ctttttgaca
     1861 tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg cgcaaatacg
     1921 catactgtta tctggctttt agtaagccgg atccacgcgt ttacgccccg ccctgccact
     1981 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     2041 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     2101 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     2161 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     2221 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     2281 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     2341 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     2401 ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     2461 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     2521 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     2581 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     2641 aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa gcataaagtg
     2701 taaagcctgg ggtgcctaat gcggccgcca tagtgactgg atatgttgtg ttttacagta
     2761 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta
     2821 cgtttctcgt tcagcttttt tgtacaaact tgtgatatcg aattcgctag ctacctttct
     2881 cttctttttt ggatctacct ttctcttctt ttttggggat ccactagttc agctcttctg
     2941 cataataact atctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     3001 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3061 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3121 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3181 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3241 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3301 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3361 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3421 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3481 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3541 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3601 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3661 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3721 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3781 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3841 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3901 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3961 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4021 cttttgcaaa aagctcccgg gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4501 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4621 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5161 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5461 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5761 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5821 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
