LOCUS       sequence-67100-e        5828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FX cloning E.coli expression vector with araBAD promoter N-terminal 
            Avi-tag, C-terminal 3C protease cleavage site and 10x His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5828)
  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 5828)
  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: pBXNAC3H
FEATURES             Location/Qualifiers
     source          1..5828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             325..369
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(418..723)
                     /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(520..539)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1065..1724)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1639..1658
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1778..1784,1785..1802,1803..1808))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1833..1957)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1976..1996)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2000..2020)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2047..2070
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2071..2100
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2111..2140
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2156..2173
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2229..2246)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2399..2485
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2577..2604
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2623..2714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2715..2783,2784..3575)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2933..2952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3746..4334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4235..4254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4488..4505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4520..4660
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4586..4605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4746..4768
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4924..5802)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggcact
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggctggtctg aacgatatct tcgaagctca gaaaatcgaa
      361 tggcacgaaa gttgaagagc gacctgcaga ctggctgtgt ataagggagc ctgacattta
      421 tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc
      481 agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg
      541 ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag
      601 cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact
      661 ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg
      721 catttcacca gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga
      781 cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt
      841 cattctgcat ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa
      901 ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt
      961 gacatacttc gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa
     1021 tacgcatact gttatctggc ttttagtaag ccggatccac gcgtttacgc cccgccctgc
     1081 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     1141 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     1201 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     1261 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     1321 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     1381 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     1441 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     1501 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     1561 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     1621 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     1681 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     1741 cctgaaaatc tcgacggatc ctaactcaaa atccacacat tatacgagcc ggaagcataa
     1801 agtgtaaagc ctggggtgcc taatgcggcc gccatagtga ctggatatgt tgtgttttac
     1861 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     1921 tttacgtttc tcgttcagct tttttgtaca aacttgtgat atcgaattcg ctagctacct
     1981 ttctcttctt ttttggatct acctttctct tcttttttgg ggatccacta gttcagctct
     2041 tctgcattag aagttttgtt tcaaggtcca caccatcatc atcaccatca tcatcatcat
     2101 taataatcta gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca
     2161 tcatcatcat cattgagttt aaacggtctc cagcttggct gttttggcgg atgagagaag
     2221 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
     2281 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
     2341 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
     2401 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     2461 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     2521 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     2581 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
     2641 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2701 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2761 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2821 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2881 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2941 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3001 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3061 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3121 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3181 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3241 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3301 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3361 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3421 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3481 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3541 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3601 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3661 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3721 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3781 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3841 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3901 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3961 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4021 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4081 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4141 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4201 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4261 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4321 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4381 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4441 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4501 gcgaggaagc ggaagtgccc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4561 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     4621 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     4681 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4741 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     4801 gcagatcaat tcgcgcgcga aggcgaagcg gcatgcataa tgtgcctgtc aaatggacga
     4861 agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct gattcgttac
     4921 caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc
     4981 gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa
     5041 accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc
     5101 ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
     5161 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
     5221 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
     5281 cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag
     5341 atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc
     5401 aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc
     5461 aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca
     5521 ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg
     5581 gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc
     5641 ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa
     5701 aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa
     5761 cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc
     5821 attcagag
//
