LOCUS       Exported                5779 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  FX cloning E.coli expression vector with araBAD promoter and 
            N-terminal 10x His tag and 3C protease cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    pBXNH3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5779)
  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 5779)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5779
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             322..351
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             352..375
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(424..729)
                     /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(526..545)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1071..1730)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1645..1664
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1784..1814)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1839..1963)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1982..2002)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2006..2026)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2062..2091
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2107..2124
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2180..2197)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2350..2436
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2528..2555
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2574..2665
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2666..3526
                     /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(2884..2903)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3697..4285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4186..4205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4439..4456
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4471..4611
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4537..4556)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4697..4719
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4875..5753)
                     /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 gcaccatcat catcaccatc atcatcatca tttagaagtt
      361 ttgtttcaag gtccaagtag aagagcgagc tgcagactgg ctgtgtataa gggagcctga
      421 catttatatt ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct
      481 gagatcagcc acttcttccc cgataacgga gaccggcaca ctggccatat cggtggtcat
      541 catgcgccag ctttcatccc cgatatgcac caccgggtaa agttcacggg agactttatc
      601 tgacagcaga cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat
      661 atcactctgt acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt
      721 aaactgcatt tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt tcaataaacc
      781 gggcgacctc agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac
      841 gggcttcatt ctgcatggtt gtgcttacca gaccggagat attgacatca tatatgcctt
      901 gagcaactga tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct
      961 ctttttgaca tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg
     1021 cgcaaatacg catactgtta tctggctttt agtaagccgg atccacgcgt ttacgccccg
     1081 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     1141 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     1201 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     1261 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     1321 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     1381 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     1441 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     1501 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     1561 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     1621 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     1681 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     1741 ttagctcctg aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa
     1801 gcataaagtg taaagcctgg ggtgcctaat gcggccgcca tagtgactgg atatgttgtg
     1861 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     1921 tatcatttta cgtttctcgt tcagcttttt tgtacaaact tgtgatatcg aattcgctag
     1981 ctacctttct cttctttttt ggatctacct ttctcttctt ttttggggat ccactagttc
     2041 agctcttctg cataataact agaacaaaaa ctcatctcag aagaggatct gaatagcgcc
     2101 gtcgaccatc atcatcatca tcattgagtt taaacggtct ccagcttggc tgttttggcg
     2161 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     2221 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     2281 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     2341 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2401 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     2461 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     2521 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     2581 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2641 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2701 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2761 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2821 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2881 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     2941 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3001 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3061 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3121 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3181 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3241 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3301 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3361 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3421 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3481 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3541 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3601 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3661 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3721 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3781 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3841 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3901 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3961 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4021 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4081 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4141 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4201 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4261 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4321 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4381 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4441 cgagtcagtg agcgaggaag cggaagtgcg cctgatgcgg tattttctcc ttacgcatct
     4501 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     4561 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4621 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4681 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4741 cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt
     4801 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     4861 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     4921 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
     4981 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     5041 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     5101 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     5161 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     5221 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     5281 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     5341 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     5401 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     5461 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     5521 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     5581 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     5641 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     5701 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     5761 atactcccgc cattcagag
//
