LOCUS       sequence-67094-e        5780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FX cloning E.coli expression vector with araBAD promoter and 
            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 5780)
  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 5780)
  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: pBXC3H
FEATURES             Location/Qualifiers
     source          1..5780
                     /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..2022
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2023..2052
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2063..2092
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2108..2125
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2181..2198)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2351..2437
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2529..2556
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2575..2666
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2667..2735,2736..3527)
                     /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(2885..2904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3698..4286
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4187..4206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4440..4457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4472..4612
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4538..4557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4698..4720
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4876..5754)
                     /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 tagttcagct cttctgcatt agaagttttg tttcaaggtc cacaccatca tcatcaccat
     2041 catcatcatc attaataatc tagaacaaaa actcatctca gaagaggatc tgaatagcgc
     2101 cgtcgaccat catcatcatc atcattgagt ttaaacggtc tccagcttgg ctgttttggc
     2161 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     2221 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     2281 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     2341 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     2401 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     2461 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     2521 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt
     2581 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2641 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2701 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2761 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2821 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2881 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     2941 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3001 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3061 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3121 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3181 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3241 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3301 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3361 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3421 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3481 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3541 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3601 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3661 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3721 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3781 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3841 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3901 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3961 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4021 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4081 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4141 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4201 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4261 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4321 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4381 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4441 gcgagtcagt gagcgaggaa gcggaagtgc ccctgatgcg gtattttctc cttacgcatc
     4501 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     4561 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     4621 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4681 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4741 acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat aatgtgcctg
     4801 tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt
     4861 ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt cttcacaacc
     4921 ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag
     4981 ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa
     5041 aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa
     5101 ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct
     5161 ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac
     5221 ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa
     5281 ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt
     5341 aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa
     5401 ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg
     5461 aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat
     5521 ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt
     5581 tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg
     5641 tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag
     5701 atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt
     5761 catactcccg ccattcagag
//
