LOCUS       sequence-67098-e        5825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FX cloning E.coli expression vector with araBAD promoter and 
            C-terminal Avi-tag, 3C protease site and 10x His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5825)
  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 5825)
  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: pBXCA3H
FEATURES             Location/Qualifiers
     source          1..5825
                     /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             complement(370..675)
                     /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(472..491)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1017..1676)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1591..1610
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1730..1736,1737..1754,1755..1760))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1785..1909)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1928..1948)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1952..1972)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1999..2043
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2044..2067
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2068..2097
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2108..2137
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2153..2170
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2226..2243)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2396..2482
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2574..2601
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2620..2711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2712..2780,2781..3572)
                     /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(2930..2949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3743..4331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4232..4251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4485..4502
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4517..4657
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4583..4602)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4743..4765
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4921..5799)
                     /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 gagttgaaga gcgacctgca gactggctgt gtataaggga
      361 gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg
      421 gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg ccatatcggt
      481 ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggagac
      541 tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc
      601 aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta
      661 aaccttaaac tgcatttcac cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa
      721 taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca
      781 gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata
      841 tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc
      901 atacctcttt ttgacatact tcgggtatac atatcagtat atattcttat accgcaaaaa
      961 tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc acgcgtttac
     1021 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     1081 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     1141 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     1201 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     1261 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     1321 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     1381 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     1441 tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     1501 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     1561 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     1621 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     1681 gcttccttag ctcctgaaaa tctcgacgga tcctaactca aaatccacac attatacgag
     1741 ccggaagcat aaagtgtaaa gcctggggtg cctaatgcgg ccgccatagt gactggatat
     1801 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
     1861 tatttatatc attttacgtt tctcgttcag cttttttgta caaacttgtg atatcgaatt
     1921 cgctagctac ctttctcttc ttttttggat ctacctttct cttctttttt ggggatccac
     1981 tagttttgct cttctgcagg tctgaacgat atcttcgaag ctcagaaaat cgaatggcac
     2041 gaattagaag ttttgtttca aggtccacac catcatcatc accatcatca tcatcattaa
     2101 taatctagaa caaaaactca tctcagaaga ggatctgaat agcgccgtcg accatcatca
     2161 tcatcatcat tgagtttaaa cggtctccag cttggctgtt ttggcggatg agagaagatt
     2221 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
     2281 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     2341 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     2401 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     2461 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     2521 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     2581 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
     2641 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2701 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2761 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2821 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2881 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2941 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3001 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3061 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3121 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3181 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3241 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3301 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3361 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3421 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3481 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3541 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3601 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3661 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3721 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3781 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3841 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4201 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4261 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4321 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4381 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4441 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4501 aggaagcgga agtgcccctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4561 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4621 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4681 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4741 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca
     4801 gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc
     4861 agggattctg caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa
     4921 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
     4981 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
     5041 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
     5101 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
     5161 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
     5221 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
     5281 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
     5341 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
     5401 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
     5461 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
     5521 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
     5581 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
     5641 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
     5701 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
     5761 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
     5821 cagag
//
